Objective To investigate the change of surface molecules and cytokine expressions of dendritic cells (DC) stimulated with recombinant proteins Com1 and heat shock protein B (HspB) of Coxiella burnetii (Cb).MethodsThe DC cultured for 5 d were stimulated with 20 μg/mL recombinant protein Com1,20μg/mL HspB or 6 μg/mL E.coli lipopolysaccharide (LPS).Twentyfour hours later,the surface mature marker,CD83,and activation-associated markers of T lymphocytes,CD58,CD54,CD40,CD80 and CD86,and expression levels of interleukin (IL)-10 and IL-12 of DC were detected by fluorescence activated cell sorter (FACS).Multiple comparisons were performed by using Student-Newman-Keuls test.ResultsCom1 could induce DC maturation efficiently in vitro.Human monocyte-derived DC exhibited significantly higher expression levels of surface molecules including CD83,CD54,CD58,CD80,CD86,and CD40,which were all ＞80％.CD83 expression induced by Com1 was similar with that induced by 1E.coli LPS,while CD54 and CD86 expressions were slightly higher than those induced by E.coli LPS,and expressions of other molecules were significantly higher than those induced by E.coli LPS (all P＜0.05).After Com1 stimulation,intracellular IL-12 level increased to 9 ％ from zero.HspB could not induce DC maturation in vitro.The intracellular IL-10 level was 6％ after HspB stimulation.DC pulsed with Com1 and HspB exhibited intracellular IL-12 level of zero and IL-10 level of ＜2％.Conclusion Com1 but not HspB can efficiently activate DC and Com1-activated DC can drive T cells toward Th1 cell development due to a high level of IL-12 production.

Cystic echinococcosis is one of the most widespread and severe zoonotic helminthic diseases .To understand the phylogeny and genetic polymorphism Echinococcus granulosus (E .granulosus) prevailed in south region of Qinghai Prov-ince ,partial fragment of cytochrome oxidase Ⅰ (COX Ⅰ ) gene sequences were used to infer the phylogenetic relationship of 59 collected samples of E .granulosus in Qinghai Province .Total 72 sequences (13 sequences from GenBank) were aligned using CLUSTAL X ,and then ,Bayesian analyses were performed in Mrbayes-3 .1 .2 .The results revealed that Echinococcus spp .isolates did not form a monophyletic group .The most samples clustered with E . granulosus strain (G1) (AB297617) , but showed high genetic polymorphism .Another three samples clustered with E .multilocularis (AB018440) ,while they showed complex phylogenetic relationships among them ,further indicating that Echinococcus spp .isolates from Qinghai Prov-ince may has a more complex evolutionary history than expected .

Objective To understand the blood pressure variability (BPV) and the influencing factors through ambulatory blood pressure monitoring during hemodialysis (HD) in the end-stage renal disease (ESRD) patients.Method Eighty-one ESRD patients on maintenancing HD for more than three months were enrolled into the study.The patients were with properly dry body weight.The blood pressure was monitored using dynamic blood pressure monitor around the HD.BPV was estimated with the coefficient of variation (CV) and standard deviation (SD) of the systolic blood pressure (SBP-CV,SBP-SD).Patients were divided into two groups according to the mean of SBP-CV:high SBPV group and low SBPV group.The possible influencing factors such as age,dialysis duration,ultrafitration volume,ultrafiltration/body weight,therapy of antihypertensive,electrolyte,nutrition state,metabolic bone disease indexes,inflammatory state and serum lipid state were analyzed and compared between the two groups.And multivariate stepwise regression analysis was made between the SBP-CV,SBP-SD and the above observational parameters.Results The average SBP-CV of the 81 patients was (8.12± 3.16)％,SBP-SD was (11.22±4.55) mm Hg.The proportion of hypertention and hypotention in high SBPV(SBP-CV≥8.12％) group (20.0％,25.7％) was higher than that in the low SBPV(SBP-CV ＜8.12％) group (8.7％,6.5％)(P =0.009).Serum high-sensitivity c-reactive protein (hs-CRP) and alkaline phosphatase (ALP) were higher in high SBPV group than that in the low SBPV group[(7.19± 5.95) mg/L vs (3.35±2.78) mg/L,P =0.001 and (180.31±96.32) U/L vs (98.00±41.19) U/L,P =0.049].Serum creatinine and potassium were higher in the low SBPV group than that in the high SBPV group [(1015.83±276.20) μmol/L vs (893.63±216.61) μmol/L,P =0.034 and (5.27±0.78) mmol/L vs (4.80± 0.23) mmol/L,P =0.005].SBP-SD was positively correlated with hs-CRP (β =0.499,P ＜ 0.01),SBP-CV was positively correlated with hs-CRP and dialysis vintage (β =0.464 and 0.211,P ＜ 0.01 and P ＜ 0.05) by the multivariate stepwise regression analysis.Conclusions The SBP-CV during HD is 8.12％ in ESRD patients.Hypertention and hypotention are more often in the higher SBPV patients.SBPV is closely related to the serum hs-CRP.

Objective To explore the effect of electroporation mediated gene therapy on angiogene-sis of the distraction area during early mandibular distraction osteogenesis (DO). Methods Thirty-two New-Zeland rabbits were randomly divided into 4 groups: group A: recombinant plasmid pIRES-VEGF165-EGFP and electroporation; group B: recombinant plasmid pIRES-VEGF165-EGFP; group C: normal saline (NS) and electroporation and group D: control group. The rabbits were sacrificed at 1d, 3d, 7d and 14d after injection, respectively. The distraction area tissue was removed for histological examination and electron microscopy, and immunohistochemical stain for CD34 was performed to detect the microvessel density. Results Generation of vascular endothelial cells (VEC) in the group A and group B were active, and majority of VEC in groups C and D took on early change of cataplasia and apoptosis. The immunohistochemical stain for CD34 showed that it expressed weakly at the first day after transfection, and at 3,7,14 days after transfection, CD34 of VECs in the distraction area expressed positively. CD34 expression was the strongest in group A (P<0. 05), and there was significant difference among three groups and different time, respectively.Compared to each other, CD34 of VECs expressed positively with a tendency to rise in the groups A and B. But it fluctuated at the level of the expression at the first day in the groups C and D. Conclusion Electroporation-mediated transfecting recombinant plasmid could promote angiogenesis during early stage of mandibular DO. It could promote local vascular proliferation and penetration, increase the blood flow of broken ends in fractured bone. It indicates that electroporation-mediated transfecting recombinant plasmid play an important role in regulating and promoting growth and reparative process of the bone.

Objective To explore in vitro cytotoxic activities of DCIKs against hepatocarcinoma cells by co-culturing cytokine-induced killer cells (CIKs) with dendritic cells (DCs) derived from peripheral blood of patients with hepatocellular carcinoma (HCC).Methods Peripheral blood mononuclear cells (PBMCs) were isolated from 23 patients with HCC and cultured with cytokines to induce DCs and CIKs.DCIKs were induced by co-culturing CIKs with DCs.After 14 days of co-culture,the phenotypes of DCIKs and CIKs were analyzed by flow cytometry,and their in vitro cytotoxic activities against SMCC-7721 and HepG2 hepatocarcinoma cells were measured by MTT assay.Levels of IL-12,IL-4 and IFN-γ in the supernatants of cell culture were detected by enzyme-linked immunosorbent assay (ELISA).Results High expressions of CD3+CD8+ and CD3+CD56+ were observed on DCIKs.The percentages of effector cells,cytotoxic activity and cytokine secretion were all significantly increased with DCIKs as compared with those CIKs without DC co-culture (P＜0.05).Conclusion Co-culture of CIKs with DCs can enhance the differentiation of effector cells and the cytolytic activities of CIKs against hepatocarcinoma cells in vitro.Immunotherapy with DCIKs may be a promising strategy for the treatment of patients with HCC.

[ ABSTRACT] AIM:To explore the mechanisms of fluctuant high blood glucose-induced apoptosis of hepatocytes. METHODS:SD rats were randomly divided into normal control group ( N) , stable high blood glucose group ( S) , fluctu-ant high blood glucose group ( F) and insulin group ( I) .Diabetic rats were induced by intraperitoneal injection of strepto-zotocin (65 mg/kg) , and the fluctuant high blood glucose animal model was induced by intraperitoneal injection of ordinary insulin and glucose at different time points every day.The blood glucose fluctuation patterns of the animals in F group with-in 12 weeks were similar every day and no significant difference of the HbA1c concentration was observed compared with S group, indicating that the fluctuant hyperglycemia was successfully established in F group.The activity of superoxide dis-mutase ( SOD) and glutathione peroxidase ( GSH-Px) , and the content of malondialdehyde ( MDA) and nitric oxide ( NO) in the homogenate of the liver tissues were detected by colorimetry.The mRNA and protein levels of JNK, p-JNK, Bax and Bcl-2 were examined by RT-PCR and Western blot.RESULTS:After 12 weeks, the increases in the intakes of food and water, the urine output, and the abnormal liver function were observed in S group, I group and F group.Compared with N group, the MDA level was increased, the content of NO and the activity of SOD and GSH-Px were decreased, and up-regu-lation of JNK mRNA and p-JNK and Bax proteins, and down-regulation of Bcl-2 were also found in S group, I group and F group.The above effects were more obviously showed in F group.CONCLUSION:Oxidative stress activates JNK-MAPK signaling pathway, which is involved in fluctuant high glucose-induced apoptosis of hepatocytes.

Objective To compare the efficacy and safety of levofloxacin 750 mg for 5 days versus 500 mg for 7‐14 days intravenous (IV ) infusion in the treatment of community‐acquired pneumonia (CAP ) . Methods This study was a multi‐center , randomized , open‐label , non‐inferiority , controlled clinical trial .The CAP patients were randomized to receive levofloxacin 750 mg IV daily for 5 days or levofloxacin 500 mg IV daily for 7‐14 days .The clinical symptoms , laboratory tests , imaging results and microbiology data were collected and compared between the two treatment groups in terms of efficacy and safety .Results A total of 241 patients were enrolled in this clinical trial from 10 study centers .Among these patients ,223 were eligible for full analysis set (FAS) analysis ,including 111 in 750 mg group and 112 in 500 mg group .Of the 223 patients in FAS ,211 were eligible for per‐protocol set (PPS) analysis ,including 107 in 750 mg group and 104 in 500 mg group .Two hundred and forty‐one patients were included in safety set (SS) ,including 121 patients in 750 mg group and 120 in 500 mg group .The median treatment duration was 5 .0 days in 750 mg and 9 .0 days in 500 mg group .The median total dose was 3 750 mg in 750 mg group and 4 500 mg in 500 mg group .The overall efficacy rate was 86 .2% in 750 mg group and 84 .7% in 500 mg group in terms of FAS at visit 4 ,which suggested that the efficacy of 750 mg group was non‐inferior to 500 mg group .Of the 111 FAS patients in 750 mg group ,40 were bacteriological evaluable ,and 41 strains of pathogens were isolated .Forty‐nine of the 112 FAS patients in 500 mg group were bacteriological evaluable ,and 51 bacterial strains were obtained .The bacterial eradication rate was 100% in both groups .The clinical treatment efficacy rate for atypical pathogens was 100% in both groups .In 750 mg group ,the most common clinical adverse drug reactions (ADRs) were injection site adverse reactions including injection site pruritus ,pain and hyperemia .The other common ADRs were insomnia ,nausea ,skin rash .The most common drug‐related laboratory abnormalities were neutrophil percentage decreased , decreased white blood cell (WBC ) count , alanine aminotransferase (ALT) and aspartate aminotransferase (AST) elevation .Most of the ADRs were mild in severity and well‐tolerated .The safety profile of the two treatments was comparable in terms of the drug‐related treatment discontinuation and the incidence of ADRs .Conclusions The short‐course regimen of levofloxacin 750 mg IV for 5 days is at least as effective and well tolerated as the long‐course regimen of 500 mg IV for 7‐14 days in treatment of CAP .

Objective To investigate the effect of antithrombotic strategies on cerebrovascular events in aged patients with nonvalvular artial fibrillation(nvAF).Methods The clinic data of 101 patients aged 80 years and over with nvAF admitted from January 2005 to June 2016 were retrospectively analyzed.The stroke and transient ischemic attack (TIA) occurred in 29 patients (28.7％,stroke/TIA group) and the remaining 72 patients were assigned in non-stroke/TIA group.The antithrombotic schemes and the antithrombotic drug-related bleeding were compared between two groups.Results Among 101 patients,there were 70 cases receiving antiplatelet therapy,19 cases receiving anticoagulation therapy and 12 cases receiving no antithrombotic therapy.The nvAF history was longer in the stroke/TIA group than that in non-stroke/TIA group [15.0(8.0,17.5) vs.6.0(3.0,12.8),Z=-3.645,P＜0.01];the proportion of anticoagulation therapy was higher in non-stroke/TIA group than that in stroke/TIA group (18/72 vs.1/29,x2=5.778,P＜0.01).The occurrence of medication-related bleeding was 15.6％(14/90) in patients with antiplatelet therapy and 12.0％(3/25) in patients with anticoagulation therapy (x2=0.196,P=0.658).There was no significant difference in the first time of stroke/TIA attack among patients with different antithrombotic schemes(x2=0.859,P=0.651).Conclusion The aged patients with nvAF are in a high risk of thromboembolism,but the proportion of using antithrombotic therapy is low.Anticoagulation therapy has a protective effect against the occurrence of stoke/TIA without increasing the risk of bleeding,which makes anticoagulation therapy advisable in the elderly patients.

Objective: To describe the prevalence and relationship between moderate to vigorous physical activity(MVPA), screen time (ST) and depressive symptoms among middle school students in Guangzhou, and to provide a reference for adolescent depression prevention and control. Methods: Using stratified cluster random sampling method, a total of 2 140 students were selected from one urban area and one suburban country for MVPA, ST and depressive symptoms investigation. Logistic regression was used to analyze relations among MVPA, ST and their combined patterns for depressive symptoms. Results: A total of 89.72% students failed to meet the MVPA recommendations, girls (95.52%) were higher than boys(84.38%). Totally 67.38% of the students failed to meet the ST recommendations, boys (69.75%) were higher than girls(64.81%). The prevalence of depressive symptoms was 27.01%, girls (31.58%) higher than boys(22.80%). MVPA, ST failure compliance rate and depression symptoms detection rate of vocational high school and ordinary high school were higher than junior high school ( χ 2=6.40,121.71,27.37, P ＜ 0.05 ). Logistic regression analysis showed that the OR for depressive symptoms was 1.19 ( P =0.35) in students whose MVPA time achieving 1 h<7 d in the last week, and was 1.93 ( P <0.01) in students with the ST>2 h/d independently, no significant interactions were observed. Students with both MVPA time 1 h<7 d in the last week and the ST>2 h/d were more likely to report depressive symptoms( OR=2.03, P <0.05). Conclusion Insufficient MVPA, prolonged ST and high prevalence of depressive symptoms are observed among middle school students in Guangzhou. Students with both insufficient MVPA and prolonged ST are more likely to suffer from depressive symptoms. Specific psychological care and health education regarding physical activity and screen behavior should be implemented to students with different characteristics, as well as their parents for depressive symptom prevention.

Objective: To investigate the general situation of scoliosis and influencing factors among middle school students in Guangzhou, so as to provide evidence for behavioral intervention measures. Methods: By stratified cluster random sampling method, 2 121 students from 8 middle schools of Guangzhou were selected to conduct questionnaire survey and physical examination. Results: The detection rate of scoliosis among middle school students in Guangzhou was 8.20%, girls (9.50%)＞boys (7.00%), urban area (10.45%)＞suburb area (4.77%), senior high school (10.08%)＞junior school (6.39%) ( P ＜0.05). Among the most of the 174 positive students were found to have large thoracic curve, right lateral bending, being girls with medium scoliosis. Multivariate Logistic regression analysis showed that students in urban area ( OR=2.56, 95%CI =1.71-3.82), with mobile electronic devices usage time≥3 h/d( OR=1.59, 95%CI =1.12-2.27), prolonged near vision work ≥1 h ( OR=1.40, 95%CI =1.00-1.95), outdoor activity time＜2 h/d( OR=1.82, 95%CI =1.24-2.67) had a higher detection rate of scoliosis ( P <0.05). Conclusion The detection rate of scoliosis among middle school students in Guangzhou is much higher, which might be related to district, mobile electronic devices usage, prolonged near vision work and insufficient outdoor activity. Health education regarding spinal knowledge should be strengthened.