Objective:Normal murine DCs were pulsed with complex of tumor antigen from elemene-combo tumor cell vaccine-heat shock protein 70 of BCG (H TA-HSP70 BCG).Their proliferation and antigen presenting function were evaluated.Methods:The dendritic cells(DCs)were cultured in complete media containing GM-CSF and IL-4 and pulsed with H TA-HSP70 BCG,H TA or HSP70 BCG.Their proliferation and stimulating effects on spleen nonadherent cells were evaluated with MTT assay.Their capability of endocytosing FITC labeled dextran was assayed with FACscan,morphological changes of DC were observed in electron microscope.Results:Proliferation index of DCs pulsed with H TA-HSP70 BCG was 2.107?0.013,proliferation index of DCs pulsed with H TA-HSP70 BCG mixed with normal nonadherent spleen cell was 1.927?0.073.The percent of DCs endocytosed FITC labeled dextran was 58.61%.Above changes were more significant than those of DCs pulsed with H TA or HPS79 BCG.Conclusion:H TA-HPS70 BCG had more potent activity to pulse DC and strengthen antigen presenting of DC.

Contraindications ; Humans ; Laryngeal Neoplasms ; classification ; pathology ; surgery ; Laryngectomy ; methods ; Neoplasm Staging

Objective To describe the distribution of body mass index (BMI) and evaluate the prevalence of overweight and obesity among first to sixth graders in 8 Chinese provinces.Methods Data came from the baseline survey of the Chinese Center for Disease Control and Prevention (CDC) funded project Study on Intervention Models for Main Chronic Diseases among children in China.A stratified random cluster sampling strategy was used to select study participants in 8 provinces,including Inner Mongolia,Jiangsu,Anhui,Shandong,Hunan,Guangxi,Gansu provinces,and Chongqing,with diverse geographical and economic level distributions in China.Counties in each province were stratified into three strata according to economic level,and one county was selected systematically from each stratum.In each county,one urban and one rural elementary school were randomly selected ; and in each school,2-3 classes were randomly selected from each of 1-6 grades.All students in these selected classes were invited for participation.Physical examination was performed by trained local CDC staff using standardized instruments to measure height and weight.Overweight and obesity were diagnosed based on the BMI classification standards for Chinese children and adolescents by Working Group for Obesity in China (WGOC).Results A total of 26558(14225 boy,12333 girl) students aged 6-12 years eventually participated in 8 provinces.The average age was 9.3 ±1.8 years old.Mean BMI among boys (17.0 kg/m2) was higher than that among girls (16.4 kg/m2,t =19.60,P＜0.01).There were statistically significant differences in mean BMI of each age group of boys and 6-11 age groups of girls between urban and rural regions and among those with three economic levels (P＜0.05).The 85th and 95th percentiles of BMI among boys aged 7-11 years in urban region were higher than BMI reference in the standards from WGOC.Lower percentiles were observed among girls aged 7-12 years in rural region.The prevalence of overweight and obesity were 9.3％ and 6.5％ among schoolchildren aged 7-12 years.The prevalence of overweight among boys (10.9％) was higher than that among girls(7.6％,x2 =88.89,P＜0.01) ; while prevalence of obesity were 8.0％ among boys and4.7％ among girls,respectively (x2 =107.17,P＜0.01).The prevalence of overweight was 11.3％ in urban and 7.4％ in rural children(x2 =131.94,P＜0.01).The prevalence of obesity was 8.7％ in urban and 4.3％ in rural children(x2 =188.99,P＜0.01).The prevalences of overweight were 10.3％,8.6％ and 9.0％ in individuals with high,medium,and low economic levels,respectively(x2 =10.76,P＜0.01) ; while the prevalences of obesity were 8.2％,5.6％,and 5.4％,respectively (x2 =51.71,P ＜ 0.01).Conclusions The overall prevalence of overweight and obesity in Chinese school children was high.The prevalence in urban region with high economic level was significantly higher than that in rural region with middle and low economic level.The nutritional and behavioral intervention programs are needed to control the rising trend of obesity in children.

In this experiment, Hca-F or L615 elemene combo-TCV (H-TCV and L-TCV), H-TCV lysates, corynebacterium parvum (CP) were used to immunize 615 and Balb/c inbred mice, and their splenic DCs were prepared and pulsed in vitro with tumor cell lysates (H or L) and TCV lysates (TH or TL). The capacities of DCs to stimulate the proliferation of syngeneic nonadherent spleenic cells were tested with MTT assay. The results showed that the splenic DCs from normal mice in vitro pulsed with TH or TL could induced syngeneic noradherent splenic cells to proliferate (P＜0.01), while the H or L pulsed DCs could not. The splenic DCs from H-TCV or L-TCV or TH immunized mice re-pulsed in vitro with TH or TL exhibited stronger stimulating effects than the DCs from normal mice pulsed in vitro for the firth time pulsed with TH or TL (P＜0.01 or P＜0.05); The capacities of DCs to induce proliferation of syngeneic nonadherent splenic cells could be further enhanced by CP immunization, especially when were pulsed with TH (P＜0.01). Normal inbred 615 mice were transferred with DCs pulsed with lysates of elemene TCV (TDCs) or pulsed with lysates of Hca-F tumor cells (HDCs) on day 7 before challenged with lethal dose of live Hca-F cells, significant adoptive immunoprotective effects were seen, with 61.6% tumor inhibition rate and 25% survival in TDC adoptive transfer group. This study indicated that DCs might play a role in the mechanisms of active immunization and pulsing DCs with lysate of elemene combo TCV and isolating DCs from elemene combo TCV immunized mice were useful methods for DCs vaccine preparation.

Elemene is a new anticancer drug isolated from a Chinese traditional medicine Curcuma aromatica. In previous work, we discovered that tumor cell vaccine (TCV) treated with oleum Curcuma aromatica or elemene could induce significant immunoprophylactic effect against a variety of aminal tumor strains and the method of preparation of elemene combo-TCV(EC-TCV) already got China's inventive patent. In this paper we further studied the active immunotherapeutic effect and the possible cellular/molecular mechanisms of EC-TCV immunization. The results were as follows:(1) EC-TCV immunization showed significant therapeutic effects (P＜0.05) against murine Ca761 syngeneic mammary carcinoma (H-2k) and HCa-F allogeneic hepatic carcinoma (H-2-) models; (2) The spleen cells of Hca-F EC-TCV immunized mice displayed higher cytotoxicity and IL-12 level while the secretion of IL-10 was decreased (P＜0.05); (3) Similar to heat shock, elemene(E), mitomycin C(MMC) and glutaraldehyde (G) could act alone as stressor, and induce significant changes of the expression of membrane heat shock proteins(HSP70 or/and HSP90) on L615 leukemia and HCa-F hepatoma cells and the EC-TCVs (E+MMC+G treated in combination) showed the highest level of membrane HSPs expression (P＜0.05 or P＜0.01 );(4) The HSP70-peptide complex isolated from HCa-F EC-TCV through ADP-agarose affinity chromatographic system could induce active immunoprotection against lethal dose challenge of HCa-F hepatic cancer cell but could not protect against the cross challenge of lethal dose of L615 leukemia. The results indicated that the immunoprotective effect of EC-TCV was in some extent tumor-specific, MHC-nonrestricted, and HSPs might play an important role in its molecular mechanisms.

Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time,and cell apoptosis was induced.Cell apoptotic rates were analysed by flow cytometry,and sensitivity of MML-1 cells to apoptosis was determined.The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3.Cyclin A,B_1 and E were selected as cell cycle markers for S,G_2/M and G_1 phase of MML-1 cells,and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h.After induction by Fas for 4 h,the active form of caspase-3 was mainly expressed in cells of G_1 phase,while rarely in cells of S and G_2/M phase.Cells with negative cyclin A and B_1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent.Cells entering into late G_1 and early S phase first express active form of caspase-3,and their sensitivity to Fas-mediated apoptosis is the highest.

AIM:To investigate the cause of blindness, except those caused by cataract, in Haimen city.METHODS:According to the WHO`s criteria of blindness, the blindness level was decided through ophthalmic tests by associate chief or chief ophthalmologists who were trained especially for disability evaluation.The analysis of the the leading cause were taken too.RESULTS:Totally 3 266 persons were blindness, in which 2 118 were first level blindness, 1 148 persons were second lever blindness, and 1 308 persons were male, 1958 were female.The leading cause of blindness were retina and uveitis diseases (31.58%), genetic diseases(23.47%), cornea disease(14.49%).CONCLUSION:The leading cause of blindness are retina and uveitis diseases, genetic diseases, cornea diseases in Haimen city of Jiangsu province.Early prevention and treatment should be strengthened to reduce the occurrence of blindness.

AIM: To explore the probable mechanisms of diabetes-induced arrhythmias. METHODS: Diabetes was induced in male SD rats,using a single injection of alloxan into tail vein. Untreated age-matched animals were used as controls. All animals were observed by 2,4,6 and 8 weeks,respectively. Transmembrane potentials were recorded with conventional glass microelectrodes. RESULTS: Action potential duration(APD) at all level (APD10,APD20,APD30,APD50,APD70,APD90) was significantly lengthened in right ventricular papillary muscle from week 2 of diabetes. At week 8,APD was more lengthened at any level of repolarization than that at week 2. No differences were observed in the maximum rate of depolarization(V_ max ),overshoot(OS) and action potential amplitude(APA) as well as the resting membrane potential(RP) from the 2th to 8th week of diabetes. CONCLUSION: The results indicate that prolongation of APD may be prominently responsible for the increased incidence of cardiac re-entry-arrhythmias and sudden death,especially at late stages of diabetes.

Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Thrombasthenia ; classification ; diagnosis

Objective To explore the effects of astrocytes in cervical spinal cord posterior horn on allodynia in rat models of migraine.Methods Fifty-four male SD rats were randomly divided into 9 groups (each group n =6): blank group,sham surgery group,new inflammatory soup 3 d group (IS 3 d group),new inflammatory soup 7 d group(IS 7 d group),saline group,fluorocitrate (FCA)prevention group,FCA prevention control group,FCA treatment group,FCA treatment control group.Following the IS stimulation in the rat dual matter,Von-Frey hair was used to monitor the pain threshold of periorbital skin.Immunofluorescence technique was used to observe the expression of specific marker of astrocyte,glial fibrilary acidic protein (GFAP),and neuron activation marker,C-fos.Results ①Von-Frey hair study showed the thresholds of IS 3 d group and IS 7 d group were significantly decreased compared with the control group(3 d:5.3 ±0.3 vs 6.4 ±0.3,F =40.15;7 d:3.0 ±0.3 vs 6.3 ±0.2,F =382.5,both P ＜0.01).The pain thresholds of FCA prevention group and FCA treatment group both began to significantly increase at the day 4 (FCA prevention:5.5 ± 5.1 vs 5.1 ± 0.2,F =16.00 ;FCA treatment:4.3 ± 0.2 vs 3.0 ± 0.2,F =138.0,both P ＜ 0.01).②GFAP immunofluorescence showed that the mean optical density (A) values of IS 3 d group and IS 7 d group were significantly increased compared with the control group(3 d:24.5 ±4.4 vs 14.8 ± 2.5,F =32.54;7 d:38.9 ± 7.1 vs 14.6 ± 1.8,F =63.56,both P ＜ 0.01).()C-fosimmunofluorescence showed that mean A value of IS 3 d group was significantly increased compared with the control group (20.4 ± 2.3 vs 8.4 ± 1.1,F =129.0,P ＜ 0.01).The difference was not significant between the IS 7 d group and the control group.Conclusions Activated astrocytes contribute to the facial allodynia induced by chronic dural inflammation.Its inhibitor FCA could both prevent and treat the allodynia behaviour.All of these suggest that astrocytes may not only contribute to the initiation of mechanical allodynia but also the maintenance.