AIM: To construct a recombinant vector containing gloverin and express it in vitro by Rapid Translation System(RTS500). METHODS: The gloverin cDNA was amplified by PCR and inserted into the prokaryotic expression vector pIVEX2.3. The recombinant product was identified by PCR and enzyme digestion. The positive reconstructed expression plasmid pIVEX2.3-G was expressed by RTS500 in vitro. The expressed protein was identified by SDS-PAGE and western blotting. RESULTS: Positive recombinant plasmid pIVEX2.3-G was successfully constructed. Target protein of 13.8 Kda with 6-his tag was detected by SDS-PAGE and western blotting. CONCLUSION: Gloverin polypeptide is successfully expressed in inactive E.coli in vitro.

AIM: To investigate whether the bacterial DNA participates in SIRS and its possible mechanism. METHODS: Escherichia coli genomic DNA (EC DNA) was extracted and purified from Escherichia coli 25922 by alkaline lysis method. Mortality of mice challenged with EC DNA and the changes of TNF-? and IL-6 in rat serum were observed. ANA-1 cells were cultured in vitro, after the cells were stimulated by different concentrations of EC DNA and LPS, the level of TNF-? and IL-6 in supernatant were tested. Meanwhile,expression of TLR9 and TLR4 on cell surface was measured. Activation of NF-?B was also observed. RESULTS: The lethal effect of EC DNA on mice with an obvious dose-effect relationship was observed. The death happened within 24 hours. Calf thymus DNA and DNase I-treated EC DNA did not lead to mice to die. The changes of serum TNF-? and IL-6 in rats induced by EC DNA and LPS were similar, but TNF-? peak level of EC DNA group appeared 1 hour earlier than that of LPS group. In vitro, large amount of TNF-? and IL-6 were released from ANA-1 cells stimulated by EC DNA. High expression of TLR9 and TLR4 was observed on surfaces of THP-1 cells. In particularly, LPS induced strong activation of NF?B. The results suggested other pathway possibly took part in the signal transduction inducea by EC DNA. CONCLUSION: EC DNA has the abilities to lead to death of mice, and induces serum TNF-? and IL-6 level to increase in rats and ANA-1 cells to release cytokines in vitro. High expression of TLR9 and TLR4, strong activation of NF-?B may be its important molecular mechanism, but other pathway probably exists to play an important role.

AIM: The auxology-related indexes are important for tissue construction. In this article, we investigated the incidence of overweight and obesity and their effects on physical functions and qualities of Guangzhou adults in 2005. METHODS: 2 056 adults including 1 027 males and 1 029 females were randomly selected from Guangzhou in 2005 for physical examination. The anthropometric data such as body mass and height were collected to calculate body mass index (BMI). The subjects were divided according to the BMI criteria recommend by Group of China Obesity Task Force, and their physical functions and qualities were analyzed. RESULTS: In 2005, the detection rate of overweight and obesity in adult males were 28.7% and 5.2%, and in females were 18.6% and 5.3%. The values of diastole pressure and systolic pressure were increased with BMI, but the change tendency of step index and vital capacity/body mass was decreased with BMI. The rate of hypertension was increased with BMI, and male subjects had higher rate of hypertension than that of female subjects in all groups. Obese adult had the poorest values in vertical jump and sit-and-reach test. Those people with normal BMI had better balance ability than that of overweight and obese people. CONCLUSION: The rates of overweight and obesity in Guangzhou adult show a high level. The physical functions and qualities may be improved when the body mass increases or decreases in a certain range, but obesity has impact to those values.

Objective To investigate the protective effect of chloroquine on endotoxemia mice and its inhibition on the release of cytokines induced by LPS. Methods A total of 40 mice of Kunming species were randomly divided into four groups: LPS group received LPS at 10 mg/kg, chloroquine group received chloroquine at 20 mg/kg, LPS plus chloroquine group received chloroquine at 20 mg/kg first, then LPS at 10 mg/kg and control group received only 0.9%sodium chloride at 200 ?l/20 g. The mortality was observed within seven days after injection via caudal vein. ANA 1 cell lines were cultivated in vitro . After chloroquine was first added into the cells for 3 hours, the releases of TNF ? and IL 6 in the supernatants induced by different concentrations of LPS were measured. Results Chloroquine could decrease the death of mice due to endotoxin. Mortality dropped from 100% to 50% ( P

Objective To investigate the roles of CpG motifs in the release of cytokines induced by bacterial DNA. Methods Mice were sensitized with injection of D GalN (600 mg/kg) into the abdominal cavity 1 h before experiment. Mortality was observed after purified Escherichia coli DNA(EC DNA), calf thymus DNA(CT DNA), phosphorothioate backbone CpG oligodeoxy nucleotides (CpG ODN) and CG sequence inverted oligodeoxy nucleotides (non CpG ODN) were injected venously into mice. The injection dosage of DNA given was 30 mg/kg but ODN was 10 nmol/mouse. THP 1 cell lines were cultured in vitro . After the above reagents were added into the culture, TNF ?, IL 6, IL 1? levels were detected to observe the different abilities of these reagents to induce the release of cytokines. Results EC DNA and CpG ODN could induce the death of mice and the large amount of release of cytokines, but CT DNA and Non CpG ODN could not. Conclusion CpG ODN has the ability to induce macrophage to release cytokines largely. CpG motifs, which may play an important role in the release of cytokines induced by bacterial DNA, may probably be the structural basis of bacterial DNA.

Objective:To explore the effect of behavioral training of creative art therapy(CAT) on interpersonal communication in teenagers.Methods:Eighty teenagers were randomly divided into study group( n=40) and control group( n=40).The study group was received the behavioral training of CAT (12 times, each activity lasts for 120 min), and the control group did not take part in the behavioral training of CAT.The two groups were measured with self-esteem scale(SES), shyness scale(SS) and social anxiety scale for children(SASC) before and after behavior training.SPSS 17.0 software was used for statistical analysis, and t-test variance analysis and Mann-Whitney U test were used to compare the differences between groups or within groups. Results:Before and after behavioral training, changes in the SES total score (7.00(5.00, 9.75) vs 1.00(0, 2.00)), the SS total score (7.00(6.00, 12.75) vs 2.00(1.00, 3.00)), the SASC total score (5.00(2.00, 8.75) vs 0(0, 1.00)) and two factor scores (3.00(0.25, 5.00) vs 1.00(0, 1.00), 2.00(1.00, 4.00) vs 0(0, 1.00))in the study group were obviously higher than those in the control group( Z=-7.347, -7.338, -5.841, -4.122 and -5.393, all P<0.05).The score differences of the SASC total score ((7.19±3.82) vs (3.21±3.04)) and two factor scores ((4.00±2.23) vs (1.68±2.05), 3.00 (1.00, 5.00) vs 2.00 (0, 2.00)) of girls were significantly higher than those of boys in the study group ( t=-3.611, -3.398, Z=-2.111, P=0.001, 0.002, 0.035). Conclusion:Behavioral training of CAT can improve teenagers' self-esteem, relieve their shyness and social anxiety, as well as promote their interpersonal communication.

To improve human cognition of sub-health condition by analyzing current situation of sub-health researches,the authors of this article reviewed the concept of sub-health,etiological factor,epidemiology,diagnostic code and other sides.

OBJECTIVE:To investigate the feasibility,safety and efficacy of domestic small waist big edge-type occluder for patients with multiple outlets sac-type membranous ventricular septal defection (VSD),and summarize its technical problems and the choice of treatment strategies.METHODS:A total of 20 patients with sac-type membranous VSD,underwent left ventricular angiography at left anterior oblique 45°-60° plus CAOD 20°-25°;the left ventricular entrance diameters were 7-21 (10.9±5.2) mm,more than 2 outlets in the right ventricular surfaces,and the largest outlet diameters were 3-10 (4.8±2.9) mm.According to the result from transthoracic echocardiography (TTE) and angiography,the sac-bag size,shape,location,extent of tissue adhesion,and stability were determined.Different types of small waist big edge-type occluder were implanted,and the occluder diameter was 5-14 (4.6±2.8) mm.Following 15 minutes of blocking,the immediate effects of occlusion were observed through repeating left ventricuiar angiography and TTE.All patients rechecked ultrasonic cardiography and electrocardiogram at 5-7 days of hospital stay,and 1,3,6 and 12 months following surgery.All patients took aspirin tablets for 6 months.RESULTS:Of 20 patients,17 cases underwent domestic small waist big edge occluder,blocked successfully through left ventricular entrance,2 cases were successful using symmetry block,and 1 case was failed.Intraoperative occlusion did not affect the aortic valve and tricuspid valve function.There were 1 case with left bundle branch block and 1 case with right bundle branch block during the operation,and all recovered within a week by using hormone therapy.After 6 months,the cardiac sizes were reduced to different degrees.CONCLUSION:It is safe and effective to treat multiple outlets sac-shaped membranous VSD with domestic small waist big edge-type occluder.The key technology,according to the sac size,shape,firmness,outlet orientation,import size,and the size of aortic stump,is to determine the block site and to select a suitable occluder.

Objective　To investigate the role of bacterial DNA in systemic inflammatory response syndrome (SIRS). Methods　A total of 100 mice of Kunming species were divided into ten groups: E.coli DNA (30, 20, 10, 5 and 1 mg/kg ), 30 mg/kg of CT DNA, 60Co DNA, DNased DNA, organic residue of DNA extraction and sterile water control. The last two were pre-treated with D-galactoamine (600 mg/kg intra peritoneally). Animals were administratively injected via tail vein. General physical condition and the death rate of mice were observed within 48 h. Results　①Obvious lethal effect of double strand E.coli DNA on mice were observed with a dose-effect correlation, LD50=11.51 mg/kg. ②NO difference in death rate was found in the group of 30 mg/kg E.coli DNA with or without 60Co irradiation (10/10 and 8/10,P>0.05). ③No rats died in the group of DNased DNA, organic residue of DNA extraction and calf thymic DNA (0/10). Conclusion　Bacterial DNA may play an important role in the development of SIRS.

Objective To express anti-bacterial peptide gloverin in prokaryotic system and investigate its bioactivity. Methods E.coli BL21 was used to express gloverin. Western blot was used to identify the production and Tachypleus Amebocyte Lysate (TAL) was used to detect the bioactivity of the production. Results The production was identified as gloverin peptide by Western blotting, and the TAL data indicated that the production could neutralize LPS. Conclusion Anti-bacterial peptide gloverin is successfully expressed in E.coli BL21.