Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P ＜ 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P ＜ 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P ＜ 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P ＜ 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P ＜ 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P ＜ 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.

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.

The aim of the present study is to investigate the effects of adiponectin (APN) on hypoxia/reoxygenation (H/R) injury in cultured cardiomyocytes. Primary cardiomyocytes were obtained from neonatal rats by enzymatic digestion method and identified by immunofluorescent technique. Primary cells cultured for 72 h were used in experiment and divided into 5 groups randomly: Control group, H/R group, H/R+APN group, H/R+APN+adenine 9-beta-D-arabinfuranoside (AraA, AMPK inhibitor) group, and H/R+AraA group. The cardiocyte morphology and beating rate were observed under inverted microscope. The DNA ladder was examined by agarose gel electrophoresis, and the cell apoptosis was determined by flow cytometry. Moreover, the malondialchehyche (MDA) content in myocardial cells and the superoxide dismutase (SOD) activity in the supernatant were measured using kits, the fluorescence intensity of intracellular Ca2+ was observed by laser scanning confocal microscope, and the phosphorylation of AMPK was determined by Western blotting. Compared with control group, H/R group showed increased apoptotic rate, oxidative stress level, intracellular Ca2+ concentration and phosphorylation level of AMPK (P<0.05), while significant ameliorations in the above indices were seen in H/R+APN group. On the contrast, AraA attenuated the protective effect of APN and decreased the phosphorylation of AMPK. These results suggest that adiponectin can protect cardiomyocytes from H/R-induced oxidative stress and apoptosis through AMPK pathway.
AMP-Activated Protein Kinases ; metabolism ; Adiponectin ; pharmacology ; Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cardiotonic Agents ; pharmacology ; Cell Hypoxia ; Cells, Cultured ; Female ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; cytology ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects

Objective To explore the impacts of different excipients on physical and chemical properties of Acteoside solid lipid nanoparticles (Acteoside-SLN),and make experimental evidence for the study of prescription SLN.Methods Emulsification-evaporation was appropriate for the preparation of Acteoside-SLN.Single variable method was used for fumbling the effects of Compritol 888 ATO,glyceryl monostearate,soy lecithin,Myrj52 and other accessories on the physicochemical properties including nanoparticles particle size,encapsulation efficiency and characterization dispersity (PDI) of Acteoside-SLN.Transmission electron microscope was used to observe the morphology of Acteoside-SLN and the structure of Acteoside in SLN was measured by XRD.Results With the increasing of the amount of Compritol 888 ATO,the particle size of nanoparticles decreased significantly,encapsulation efficiency decreased slightly,PDI increased;With the increasing of amount of glycerol monostearate,particle size increased obviously,encapsulation efficiency decreased slightly,and PDI decreased;With the increasing of the amount of lecithin,particles size increased significantly,encapsulation efficiency decreased,and PDI decreased;With the increasing of Myrj52 amount,the nanoparticles particle size decreased,encapsulation rate and PDI increased slightly.The appearance of Acteoside-SLN was presented uniform spherically.Acteoside was wrapped in SLN in a molecular dispersion state.Conclusion Various additives have a greater impact on physicochemical properties of Acteoside-SLN,and inspiration for prescription screening of SLN is supplied by this study.

OBJECTIVETo understand the epidemic characteristics and viral antibody level among healthy people of Japanese B encephalitis (JE) in Henan province in 2010.

METHODSA total of 433 selected JE cases in Henan province in 2010 were analyzed by descriptive epidemiological method. Xinyang and Luoyang were selected as survey sites in 2010.12 administrative villages were randomly selected from both cities. As the investigation objects, 519 healthy people from the two cities were randomly selected by eight age groups:less than one year old, 1 - 2 years old, 3 - 4 years old, 5 - 6 years old, 7 - 14 years old, 15 - 19 years old, 20 - 59 years old, and above 60 years old. A total of 1008 samples of blood specimens were collected both in May and November, and JE viral antibody was detected by micro-cytopathic effect neutralization test.

RESULTSThe incidence rate of JE was 0.46/100 000 (433/94 130 434) in Henan province in 2010.97.69% (423/433) of the patients were found between July and September, and 81.06% (351/433) were distributed in Nanyang, Xinyang, Luoyang, Zhumadian and Zhoukou city. Children aged 0 to 14 years were the primarily affected group (82.22%, 356/433), the people above 15 years old accounted for 17.78% (77/433) of whole cases in Henan province, but the same group accounted for 65.79% (50/76) of whole cases in Luoyang city, which obviously higher than the percentage in Henan province (χ(2) = 79.57, P < 0.05). Most patients were scattered children in Henan province, accounting for 58.89% (255/433). In Luoyang city, most patients were peasants, accounting for 44.74% (34/76). The antibody positive rate of JE among health people above 15 years old in Luoyang city was 48.94% (46/94), which was lower than it in Xinyang city at 97.78% (88/90). The difference showed statistical significance (χ(2) = 55.42, P < 0.05). The antibody positive rate among healthy people under vaccination was 50.41% (61/121), which was obviously higher than that among people without vaccination, at 16.67% (6/36) in Luoyang city. The difference showed statistical significance (χ(2) = 12.92, P < 0.05). The antibody positive rate among healthy people under vaccination was 67.11% (51/76) in Xinyang city, which was obviously higher than that among people without vaccination, at 46.39% (45/97). The difference showed statistical significance (χ(2) = 7.40, P < 0.05).

CONCLUSIONThe incidence of JE showed seasonal and regional characteristics, there were differences among ages and occupations. The difference was consistent with the difference in viral antibody level among health people in Henan province and Luoyang city.

Adolescent ; Adult ; Antibodies, Viral ; blood ; Case-Control Studies ; Child ; Child, Preschool ; China ; epidemiology ; Encephalitis Virus, Japanese ; immunology ; Encephalitis, Japanese ; blood ; epidemiology ; immunology ; Humans ; Incidence ; Infant ; Middle Aged ; Young Adult

OBJECTIVETo evaluate the clinical application value of flexible multi-analyte profiling (xMAP) technology in detecting high-risk human papillomavirus (HR-HPV).

METHODSTotally 1 061 women, aged 21-65 years, were randomly enrolled into the study. Cervical exfoliated cells were used in xMAP technology and hybrid capture II (hc2). Pathological diagnosis was used as golden standard. Consistency of these two methods was assessed.

RESULTSThe sensitivity and specificity of xMAP technology were 80.31% and 85.83%, respectively. The positive and negative predictive values were 44.5% and 96.9%, respectively. The Kappa value for consistency between xMAP technology and hc2 was 0.58.

CONCLUSIONSThe specificity of xMAP technology is similar to hc2 test, but the sensitivity is inferior to hc2. However, these two methods show good consistency in the detection of HR-HPV.

Adult ; Aged ; Cervix Uteri ; virology ; Female ; Humans ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; virology ; Sensitivity and Specificity ; Uterine Cervical Diseases ; diagnosis ; virology ; Virology ; methods ; Young Adult

Objective To know the immunologic response at different periods with different hepatitis B vaccine(HepB)in infants.Methods From July 1 to December 31 in 2009,the local infants born from hepatitis B virus(HBV)surface antigen (HBsAg)negative mother in Ninghai were selected and received a 0.5 ml of 5 -μg HepB made by recombinant deoxyribonucleic acid (DNA)techniques in saccharomyces cerevisiae yeast (HepB -SCY)and those in Yuyao were selected and received a 0.5 ml of 10 -μg HepB made by recombinant DNA techniques in hansenula polymorpha yeast (HepB -HPY)with a 0 -1 -6 month schedule.The specimens of blood were collected at 7 months,12 months and 18 months after vaccination for testing antibody.Results At 7 months,12 months and 18 months after vaccination,the geometric mean concentration (GMC)of Anti -HBs in HepB -SCY group were 714.79 ±3.50 mIU /mL,259.89 ± 3.87 mIU /mL and 78.38 ±4.04 mIU /mL,respectively.While in HepB -HPY group were 2 420.76 ±3.51 mIU /mL, 703.55 ±3.46 mIU /mL and 171.32 ±3.76 mIU /mL,respectively.There was statistical significant difference of GMC between HepB -SCY group and HepB -HPY group(P <0.01).The percent of antibody <100 mIU /mL in HepB -SCY group were 6.39%,23.33% and 55.42%,and in HepB -HPY group,were 2.17%,5.17% and 31.95%,respectively. The difference between HepB -SCY group and HepB -HPY group had significantly statistical significance (P <0.01 ). Conclusion The immunologic response at different periods of HepB -HPY was better than that of HepB -SCY.

OBJECTIVETo report and study a case of sporadic family fatal insomnia (SFFI) on its.

METHODSInvestigate clinical characteristics and family disease history of a suspect FFI patient. His clinical characteristic was analyzed, he and his 14 family members genomic DNA was extracted by standard techniques from their and blood detected with polymerase chain reaction (PCR) method and DNA sequencing to find out his prion protein (PrP) gene mutation. The patient's CSF was detected with Western-Blot method for 14-3-3 brain protein.

RESULTSThe patient was diagnosed as an sporadic FFI by his developed sleep disturbance and changes in sleep-awake rhythm, motor abnormalities, mental disorder, dementia, autonomic dysfunction; his family history; his 14-3-3 brain protein-positive (CSF) and analysis results of his PrP gene (codon point mutation D178N and methionine homozygosity at position 129M/M). Suggesting that in the future to identify CJD and FFI patients, screening should focus on clinical symptoms and laboratory results. The PrP gene of 14 family members did not appear Mutation, and there is no person suffering from the same disease.

CONCLUSIONSThe case was diagnosed as a sporadic familial fatal insomnia. Analysis of suspicious patients' genomic DNA for PrP gene mutation might be very important for FFI diagnosis because there exist many difficulties in clinical laboratory evaluation. This patient might be the first SFFI patient reported in China and the case finding might have momentousness in clinical and basical study.

14-3-3 Proteins ; genetics ; Humans ; Insomnia, Fatal Familial ; genetics ; Male ; Middle Aged ; Mutation ; PrPSc Proteins ; genetics

Objective To analyze interspecies cross-contamination of 160 non-human cell lines.Methods One hundred and sixty common non-human cell lines were collected and their species were identified by PCR.For the suspicious cells,chromosome analysis was further used to confirm their species.Results Six in 160 non-human cell lines were cross-contaminated.A rat cell line was mixed by a human cell line,and 5 were totally cross-con-taminated,and were indentified as wrong species.Conclusions Species identification is an indispensable part of cell quality control.Each cell line should undergo a full QA(Quality Assurance)assessment before it is used for research.

Bismuth modified boron doped diamond (BDD) film electrode was employed for simultaneous determination of trace ZnⅡ,CdⅡand PbⅡby anodic stripping voltammetry.BiⅢwas simultaneously in-situ deposited on bismuth modified boron doped diamond electrode with ZnⅡ,CdⅡ and PbⅡ by pre-concentration.In the presence of BiⅢ,the sensitivity for determination of ZnⅡ,CdⅡ and PbⅡ was remarkably enhanced.Influence factors such as bismuth concentration,boron doped concentrations of BDD electrode,pH,preconcentration potential were investigated and optimized.Under the optimal conditions,the stripping peak currents increased linearly with the increasing concentration of ZnⅡ,CdⅡ and PbⅡ in the range of 10-300 μg/L.The limit of detection was 0.56 μg/L for ZnⅡ,0.32 μg/L for CdⅡand 0.75 μg/L for PbⅡ (S/N=3),respectively.The interference experiments showed that common ions had little influence on the determination except CuⅡ.In addition,the developed electrode displayed a good repeatability.The method was successfully applied to determination of ZnⅡ,CdⅡ and PbⅡ in real water samples with the standard addition recoveries of 92.0%-114.0%.