OBJECTIVE: To explore the relationship between the expression changes of cytokines, interleukin-6 (IL-6), interleukin-8(IL-8), and tumor necrosis factor-alpha (TNF-alpha), and the wound time following explosive injury to rabbit's chest. METHODS: The rabbit's model of explosive injury was established. The expression levels of IL-6, IL-8 and TNF-alpha in the plasma were detected by ELISA method at different wound time (0.5-12h). RESULTS: The level of IL-6 increased at 3h after wounding and reached peak at 6 h. The level of IL-8 increased at 1 h and reached peak at 6 h. The level of TNF-alpha increased at 0.5 h and reached peak at 3 h. CONCLUSION IL-6, IL-8 and TNF-alpha have a time-related expression after explosive injury.
Animals ; Enzyme-Linked Immunosorbent Assay ; Explosions ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Rabbits ; Thoracic Injuries/metabolism* ; Tumor Necrosis Factor-alpha/metabolism*

Objective To explore the expression and the clinical significance of neural cadherin (N-cadherin) in perihilar cholangiocarcinoma. Methods The expression of N-cadherin was detected by immunohistochemical method in 26 patients with perihilar cholangiocarcinoma and 5 samples of normal bile duct tissues. The expression of N-cadherin was also compared by Western blotting assay between perihilar cholangiocarcinoma and paraneoplastic tissues. Results The im-munohistochemical results showed that positive expression of N-cadherin was observed in 14 of 26 patients with perihilar cholangiocarcinoma and in 6 of 26 paraneoplastic samples (53.84%vs 23.08%, P<0.05). There was 0 N-cadherin expres-sion in normal bile duct tissues. The positive rate of N-cadherin expression was higher in low differentiated group than that in middle and well differentiated groups. The result of Western blotting assay showed that N-cadherin expression was signifi-cantly higher in perihilar cholangiocarcinoma than that in the paraneoplastic tissues (0.88 ± 0.12 vs 0.43 ± 0.09, P < 0.05). Conclusion N-cadherin may be involved in the occurrence and development of perihilar cholangiocarcinoma.

Objective To explore the relationship between obesity phenotypes and adipocytokines in children.Methods Based on the Beijing child and adolescent metabolic syndrome (BCAMS) study,3 508 children (1 788 boys and 1 720 girls) aged 6-18 were recruited.In this study,participants were categorized into four groups:226 cases in general obese group,192 cases in abdominal obese group,1 004 cases in combined obese group and 2 086 cases in non-obese group,according to the sex,age,specific body mass index(BMI),and waist circumference (WC) equal to or greater than the 90th percentile for age and gender of school children in Beijing in 2004.The levels of plasma insulin,serum leptin,resistin and adiponectin were measured by sensitive,specific double-antibody sandwich enzyme-linked immunosorbent assays (ELISA).Analysis of covariance,multivariate linear regression and binary logistic regression analysis were performed.Results There were highest plasma insulin and serum leptin,and lowest adiponectin levels in combined obese group than those in other obese groups and non-obese group and resistin level in abdominal obese group was highest than those in other obese groups or non-obese group.Among subjects with general obesity and conbined obesity,WC was more important factor than BMI for plasma insulin[?(WC)=0.158 P0.05].With covariates adjusted,the odds ratios(OR)and 95% confidence intervals of general obesity,abdominal obesity and combined obesity were 3.46(2.44-4.91),5.41(3.87-7.57) and 10.10(8.26-12.35) for predicting hyperinsulinemia,respectively,5.83(4.02-8.45),7.07(4.97-10.05)and 20.82(16.49-26.28) for hyperleptinaemia,respectively,1.47(1.05-2.07),2.0(1.42-2.80) and 2.66(2.23-3.18) for hypoadiponectinaemia,respectively.Serum resistin was highest in abdominal obesity.Conclusion The levels of adipocytokines in children were correlated with the phenotypes of obesity,especially for abdominal obesity.

Objective To analyze the CT findings of melamine induced urinary calculi.Methods Nineteen children with a history of ingestion of melamine contaminated infant formula milk were studied, including 12 males and 7 females, age ranged from 50 days to 5 years.Results CT demonstrated renal pelvic and ureteral stones in 13 cases, with urinary obstruction in 9 of them.The size of the stones ranged from 0.3 cm × 0.3 cm to stag-horn calculus.The density of the stones measured from a low of 40-70 HU up to a high of 410 HU with an average density of 160 HU.Conclusion CT scan is an excellent modality in demonstrating urinary tract calculi caused by melamine. It is the method of choice when ultrasound examinations are equivocal.

OBJECTIVETo study the killing effects of the liposome-mediated thymidine kinase (TK)/ganciclovir (GCV) system on MG-63 osteosarcoma (OS) cells and its bystander effects.

METHODSLiposome-mediated TK gene transfected into MG-63 OS cells, the efficiency of transfection was analyzed by flow cytometry and observed under inverted fluorescence microscope. Non-transfected osteosarcoma MG-63 cells were divided into three groups,in the experimental group 1 transfected TK/GCV cells cultured in solutiona liquid mixture by supernatant by 1/10,1/7,1/5,1/2 ratio to original broth; in the experimental group 2 transfected cells cultured in solutiona liquid mixture of supernatant filtered through 0.22 microm filter by 1/10,1/7, 1/5, 1/2 ratio to original broth, in control group the transfection cells cultured in original culture solution. Cell growth inhibition rate and osteosarcoma cell sensitivity to TK/GCV system were measured by MTT assay in each group.

RESULTSThe TK gene was transfected into MG-63 OS cells successfully by liposome-mediated, flow cytometry instrument detection TK gene transfection cell transfection efficiency can reach 75.5%. Six days later the MTT assay showed that in the experimental group 1 inhibition rate of all concentration ratio of the mixed culture fluid were statistically significant as compared with the control group (P < 0.05), and in the experimental group 2 that of the 1/10 and 1/7 of concentration ratio of mixed culture medium was not statistically significant as compared with the control group (P > 0.05). TK gene transfected MG-63 cells increased with the the GCV concentration,the cell apoptosis rate increased.

CONCLUSIONThe experiment demonstrated that the MG-63 OS cells are sensitive to the liposome-mediated TK/GCV system and bystander effects are significant.

Apoptosis ; drug effects ; Bone Neoplasms ; enzymology ; genetics ; physiopathology ; Bystander Effect ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Ganciclovir ; toxicity ; Humans ; Osteosarcoma ; enzymology ; genetics ; physiopathology ; Thymidine Kinase ; genetics ; metabolism ; toxicity

ObjectiveTo evaluate the technical feasibility of animal model of avascular necrosis of femoral head (ANFH)with transcatheter arterial embolization (TAE).MethodsTwenty experimental pigs were randomly divided into experimental group and control group (each n= 10).Experimental group:A 5F Cobra catheter was inserted into left femoral artery,and the feeding arteries of femoral head were superselectively inserted.The feeding arteries were embolismed through transcatheter arterial injecting the segments of silk measuring about 500μm.Control group:The arterial embolization was not performed,and the other treatment was identical to experimental group.The articulation of hip in all pigs underwent plain X-ray examination,CT and MR scanning 2 weeks and 4 weeks after treatment,respectively.Histological examination was made in 4 weeks to evaluate volume of bone trabecula (TBV) and percentage of bone lacuna (PBL) at unit area under microscope.The data were compared between the two groups.Results In experimental group,CT and MRI showed swolling in hip soft tissue and high T1 in hip joint cavity,while no obvious abnormalities were found in plain X-ray film 2 weeks after feeding arteries were embolized.Four weeks after feeding arteries embolization,plain X-ray film,CT and MR showed typical necrosis of femoral head in the experimental group,while no obvious abnormalities were found in control group.The histology examination revealed there were obvious karyopyknosis and anachromasis in the bone cells.The quantity of bone cells decreased obviously or disappeared.PBL increased and TBV decreased significantly compared with those of control group (P＜0.05).ConclusionThe animal model of ANFH in pigs can be induced by TAE.It can preferably mimic the pathological situation of ANFH.

Objective To retrospectively analyze the effect of interventional embolization for hepatocelluar carcinoma (HCC) associated with arteriovenous shunting (AVS), and to discuss the factors influencing the therapeutic results. Methods The clinical data of 62 cases with HCC associated with AVS , who were treated with interventional chemoembolization , were retrospectively analyzed. Based on the type and extent of AVS identified by angiographic manifestations, appropriate obstruction of the shunt and Lipiodol chemoembolization of HCC were conducted. The curative effect of the shunt embolization was assessed by DSA at one or two months after the treatment. The relevant factors influencing the prognosis of embolization were analyzed by using univariate and multivariate Cox regression analysis methods. Results Of the 62 patients, arterioportal shunting (APS) was detected in 44, hepatic arterio-venous shunting (HAVS) in 11, APS together with HAVS in 4, and hepatic artery-pulmonary artery shunting (HAPAS) in 3. Re-examination with DSA was carried out in 53 patients at 1 - 2 months after the treatment , which showed that the shunting disappeared in 18 cases, obvious reduction of the shunt flow was seen in 19 cases, the lesion remained stable in 9 cases and the disease became worse in 7 cases. Univariate analysis indicated that the kind of embolic material and the presence of tumor thrombus could affect the obstructive result of the shunt , while multivariate Cox regression analysis showed that portal tumoral thrombus was an independent risk factor. The embolization effect of polyvinyl alcohol (PVA) particles and Lipiodol-ethanol mixture, used as the embolic agents, was better than that of gelatin sponge particles. Conclusion To ensure a successful interventional chemoembolization for HCC combined with AVS the procedure should be individualized according to the type and extent of the arteriovenous shunting. The type of embolic materials used for embolization can affect the results to a certain degree.

Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.

Objective To knowthe effect of selenium on expression ofnitric oxide synthase(eNOS) and inducible nitric oxide synthase(iNOS) induced by fluoride in ECV-304(human umbilical vein endothelial cells).Methods The cells,ECV-304,were divided into two groups and treated as follows,one group was treated with sodium fluoride at different doses(0,400,1 000 and 2 000 ?mol/L),the other was treated with sodium fluoride at different doses(0,400,1 000 and 2 000 ?mol/L)and added sodiumselenite(100 nmol/L) respectively.24 h of treatment,The activity ofeNOS and iNOS were measured by NO and NOS assay kits and the expression of eNOS mRNA and iNOS mRNA was analyzed by RT-PCR.Results The activity of eNOS and the expression of eNOS mRNA were down-regulated by fluoride with a dose dependent manner,while the activity of iNOS and the expression of iNOS mRNA were up-regulated.Selenium showed a significant antagonism to low level fluoride treated groups.Conclusion Seleniumcan antagonize the effect offluoride on the expression ofiNOSand eNOS in the vein endothelial cells when the fluoride exposure level is not toohigh.

Objective To study the effect of different concentrations of fluoride on cultured human umbilical vein vascular endothelial cells(HUVEC). Methods Different doses of sodium fluoride (NaF) were added to HUVEC culture medium, fluoride concentrations were 0(control), 100,400,700,1000,2000 μmol/L, respectively,6 re-set hole in each group. After continuous culture for 48 h, cells and culture medium were collected. Cell morphology was studied by Wright-Giemsa staining; cells apoptosis was determined by acridine orange fluorescence staining; cell activity was measured by methyl thiazolyl tetrazolium (MTT) assay; superoxide dismutase (SOD),glutathione peroxidase(GSH-Px) activity, malonaldehyde(MDA) content, induced nitricoxide synthase(iNOS), and endothelia nitricoxide synthase(eNOS) activity in cell culture medium were determined by spectrophotometry; cell iNOS mRNA and eNOS mRNA expression were detected by RT-PCR; intercellular adhesion molecule-1 (ICAM-1)and vascular cell adhesion molecule-1 (VCAM-1) levels were detected by double antibody sandwich ELISA method.Results With increased dose of fluoride, HUVEC cells decreased, the structure changed. In 400 - 2000 μmol/L group, the SOD activity[(6.627 ± 0.213), (6.668 ± 0.152), (5.935 ± 0.122), (4.755 ± 0.182)kU/L] was lower than those of the control group[(7.457 ± 0.398)kU/L, P ＜ 0.05 or ＜ 0.01], GSH-Px activity[(481.284 ± 43.785),(492.223 ± 16.474), (382.762 ± 25.167), (293.687 ± 24.881 )kU/L] was also lower than those of the control group [(585.078 ± 47.323)kU/L, P ＜ 0.05 or ＜ 0.01], MDA level[(0.609 ± 0.011 ), (0.646 ± 0.016), (0.852 ± 0.013),(1.188 ± 0.045)nmol/L] was higher than those of the control group[(0.512 ± 0.027)nmol/L, P ＜ 0.05 or ＜ 0.01];iNOS activity[(3.604 ± 0.115), (3.615 ± 0.075), (3.848 ± 0.103), (4.275 ± 0.079)kU/L] also was higher than those of the control group[(2.798 ± 0. 136)kU/L, all P ＜ 0.01], iNOS mRNA expression increased, eNOS activity [(5.539 ± 0.079), (5.503 ± 0.064), (5.226 ± 0.142), (4.809 ± 0. 107)kU/L] decreased compared to those of control group[(5.996 ± 0.155)kU/L, P ＜ 0.05 or ＜ 0.01], eNOS mRNA expression decreased; ICAM-1 levels [(0.852 ± 0. 102), (0.886 ± 0.061 ), (0.961 ± 0.158), (1.418 ± 0. 167)μg/L] increased compared to those of the control group[(0.687 ± 0.046)μg/L, P ＜ 0.05 or ＜ 0.01], VCAM-1 levels[(2.719 ± 0.197), (2.946 ± 0.167),(3.173 ± 0.225 ), (3.613 ± 0. 153 ) μg/L] was higher than those of the control group [(2.375 ± 0.067 ) μg/L, all P ＜0.01]. Conclusions High concentrations of fluoride reduce the activity of antioxidant enzymes, which leads to metabolic disorders of nitric oxide and abnormal cytokines expression, thereby inhibiting vascular endothelial cell growth, structural change and induced apoptosis. This is an important factor in high fluoride-induced vascular endothelial injury.