Objective　Standard method of testing serum fluoride was tested and verified by certified reference material CRM of free fluo ride component in cattle serum (GBW 09143～09144).The method was suited for fluor ide research to assess its practical value.Methods　ISE-standard curve,linear relationship,linear range,sensitive and precision were disired result by CRM.400 serum of fluoride in rats were measured.Results　The linear ranges of the method were 0.02～5.0 mg/L,r=0.9999,b=57.8,and test lower limit was 0.013 mg/L.The test value was equal to standard value.Conclusions　The standard method is accurate,sensitive,convenient and small sample is used;and it is easy to spread.Thus,it has been of practical value.

ObjectiveTo investigate the cell toxicity of a novel macropores calcium phosphate cement (CPC) scaffold and its influence on cell adhesion,growth and proliferation.MethodsA novel CPC material was synthesized by means of adding mannitol porogens and applying sodium solution as the cement liquid.The cell growth and proliferation in the novel CPC material extraction was observed by CCK8 assay.Scanning electron microscopy was used to observe hole diameter of the material,cell adhesion and growth in the material.The experiment of three point bending was used to test the biomechanic performance of the CPC material.ResultsThe novel CPC material reached hole diameter value of (267.43±118.01)μm,microporosity of (66.15±6.91)％.Maximum load,flexural strength and toughness of the novel CPC material was increased about one time compared to the traditional CPC(P＜0.05).CCK8 assay showed there were no significant difference of the light absorption value of cells in the CPC extraction in the 4th,6th,8th day compare to the control group (P＞0.05).ConclusionThe novel CPC material has the strong biomechanics performance,macropores,high microporosity and excellent biocompatibility,which is promising for ideal bone tissue engineering scaffold.

Objective To investigate the distribution of water-borne fluoride and the current status of water defluoridation project by improving drinking water quality in endemic fluorosis areas in Shandong province,and to provide scientific basis for making strategies in prevention and control of the disease. Methods According to "the National Technical Scheme for Endemic Disease Control in 2004, 2005 and 2006", 113 endemic fluorosis diseased counties(cities, and districts) of the province's 17 cities were screened in order to investigate the fluoride level in drinking water in fluorosis villages, recheck the fluoride level after implementing the water defluoridation project, and investigate the current status of the water defluoridation project. The fluoride level in drinking water was determined by F-ion selective electrode. Results There were a total of 5816 water defluoridation projects in the province. Most of them were carried out by drilling a deep well to get under-ground water. The wells still in good condition were accounted for 72.80% (4234/5816). Intermittent operated wells were accounted for 3.11% (181/5816). Abandoned wells were accounted for 24.09%(1401/5816). Level of water fluoride was determined in 6940samples from fluorosis villages(villages that not carry out the water defluoridation project as well as villages carried out the water defluoridation project with abandoned wells were included) and the value that lower or equal to 1.00mg/L was determined in 2987 villages which accounted for 43.04% (2987/6940). Level of water fluoride that over 1.00 mg/L was found in 3953 villages which accounted for 56.96% (3593/6940), and the highest level of water fluoride was 11.33 mg/L. Level of water fluoride were determined in 4415 samples from water defluoridation project and the value lower or equal to 1.00 mg/L was in 2983 wells which accounted for 65.53%(2983/4415). The value over 1.00 mg/L was in 1522 wells which accounted for 34.47%(1522/4415), the highest value of water fluoride was 9.71 mg/L. Conclusions Level of water fluoride in up to 1/2 of the villages and 1/3 of the projects, is still higher than the standard in Shandong province. Nearly 1/4 of the project has been abandoned. The current situation for endemic fluorosis control is still not good in the province, countermeasures for endemic fluorosis must be carried out as soon as possible and surveillance of water defluoridation project must be strengthened.

Objective To investigate the spatial distribution and clustering areas of Budd-Chiari syndrome in Heze City,Shandong Province,and to provide epidemiological information for further exploring the etiology and related risk factors of the disease.Methods Detailed residential addresses of 342 cases of patients (residents of Heze City) with diaphragm type Budd-Chiari syndrome diagnosed between 1995 and 2004 in Heze Municipal Hospital,Heze Shan County Central Hospital,Affiliated Hospital of Xuzhou Medical College,Shandong Provincial Hospital and Beijing Xuanwu Hospital were collected.Geographic information system (GIS) was used as a platform for data management and display.The nearest neighbor index,Ripley's K(d) function,Ripley's L(d) function and the nearest neighbor clustering method were applied to detect the spatial characters of Budd-Chiari syndrome in Heze City,Shandong Province.Crimestat 3.0 was used for spatial analysis.Results The nearest neighbor distance analysis showed that the nearest neighbor index was 0.6767 (Z =-11.4387,P ＜ 0.01).That was an aggregation at the first-order spatial scale.Within the study area,the first clustering radius of Budd-Chiari syndrome was 6.66 km,and the first clustering strength was 5.40; the average radius of the strongest clustering area was 126.61 km,and the clustering strength was 12.52,while the biggest clustering radius was larger than 222 km.After corrected by population,the gathering strength was slightly higher than that before the correction.Ten first-order hot spots were formed,and 95％ confidence interval aggregation number was 7,which meant the results were statistically significant(P ＜ 0.05),main clustering areas are in Mudan District,Shan County and Juancheng.One secondorder hot spot was gathered based on the first-order hot spot.Conclusions Spatial distribution of Budd-Chiari syndrome in Heze City,Shandong Province has showed spatial aggregation and heterogeneity.This study has a great epidemiological significance for further exploring the cause of Budd-Chiari syndrome.

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 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.

ObjectiveTo look into the distribution of “iodine suitable” region in iodine-deficient areas in Shandong province and to provide a scientific basis for guiding the redesignate of iodine-deficient areas and launch scientific supply of iodine.Methods One to 3 copies of water source samples were collected in 105 existing iodine-deficient counties by village.Water iodine content was detected by arsenic-cerium catalytic spectrophotometry.The areas with water iodine content below 10 μg/L was defined as iodine-deficient areas and among 10 - 150 μg/L were “iodine suitable areas” and greater than 150 μg/L were high iodine areas.Results The research was carried out in 14 cities,105 counties,and 1337 towns.We collected 65 716 water samples.Sample recovery efficiency reached 99.8％.The median of water iodine was 5.57 μg/L.In the 1337 towns surveyed,there were 82.05％(1097/1337) of the township with water iodine median ＜ 10 μg/L,17.43％(233/1337) between 10 - 150 μg/L,and 0.52％(7/1337) ＞ 150 μg/L.Conclusions In Shandong province,the water “iodine suitable” regions are distributed scattered with considerable proportion.In iodine-deficient areas,there are areas with high water iodine,and iodine-deficient regions should be redrawn.Emphasis should be put on iodine nutritional status of residents in “high iodine and iodine suitable” regions,and iodine supplementation should be carries out scientifically.

Objective To understand the current situation of endemic fluorosis in Shandong Province,and to provide a scientific basis for development of control strategies.Methods In accordance with the requirements of the national Surveillance Scheme of Drinking-Water-Borne Endemic Fluorosis (Trial) and Shandong Provincial Surveillance Scheme of Drinking-Water-Borne Endemic Fluorosis (Trial),and to understand the progress of the province's water-improvement projects,10 counties were chosen in the province,and 10 water-improvement projects were selected in each county.Running condition of the water-improvement projects was investigated,and water fluoride content was tested.Three epidemic villages were chosen as fixed monitoring villages in each county.In each village,fluoride content in drinking water was determined,and dental fluorosis of all children aged 8 to 12 was diagnosed.Water fluoride content was determined by fluoride ion selective electrode method,and dental fluorosis of children aged 8 to 12 was diagnosed by Dean method.Results ① There were 3570 water defluoridation projects in the province,and water-improvement rate was 87.49％ (10 122/11 569).Normal operating projects accounted for 89.10％ (3181/3570),and the passing rate of water fluoride content was 73.14％ (2611/3570),with the maximum water fluoride as 9.71 mg/L.② In the 10 counties,a total of 86 waterimprovement projects were monitored.Normal operation,intermittent operation and scrapped projects accounted for 95.35％ (82/86),3.49％ (3/86) and 1.16％ (1/86),respectively,and passing rate of fluoride content in water was 50.00％ (43/86),and the maximum water fluoride was 5.32 mg/L.③ In the 27 monitored villages with improved water quality,the passing rate of fluoride content in water was 74.07％ (20/27),and the maximum water fluoride was 4.50mg/L.In the 3 monitored villages without improved water quality,villages of mean water fluoride ≤ 1.20 mg/L and ＞ 1.20 mg/L accounted for 33.33％ (1/3) and 66.67％ (2/3),respectively,and the maximum water fluoride was 1.53 mg/L.④ In the 27 monitored villages with improved water quality,the detection rate of dental fluorosis among children aged 8 to 12 was 57.65％(791/1372),and the detection rate of defect type dental fluorosis was 7.80％(107/1372); the index of dental fluorosis was 1.15.In the 3 monitored villages without improved water quality,the detection rate of dental fluorosis among children aged 8 to 12 was 50.97％ (80/155),and the detection rate of defect type dental fluorosis was 7.10％(11/155); the index of dental fluorosis was 1.16.Conclusions In Shandong Province,the running condition of water improvement project to reduce fluoride still needs to be further improved,and the water fluoride is seriously overweight and endemic fluorosis has not been effectively controlled.Preventive measures should be strengthened.

Objective To survey the highly risk iodine deficiency regions of Shandong Province and to provide reference basis for further executing urgent iodine supply. Methods The annual document of cretinism in Licheng District in Jinan City and Shouguang City, the two iodine deficiency regions, were referred. Forty children aged 8-10 years of 2 targeted schools from 3 towns out of every targeted region underwent palpation, ultrasonography and As3+-Ce4+catalyzing speetrophotometry for ultra iodine examination. Twenty women aged 18-40 years from 2 villages sampled from every targeted town received ultra iodine examination and edible salt examination of iodine with direct titration. Results No new cretinism and suspected cretinism was found since 1995 in the regions. The goiter rates of children of the two regions detected with palpation and ultrasonography were 7.5% (18/241),6.2% (15/241) and 5.0% (13/259), 1.2% (3/259), respectively. Two hundred forty and 249 urine samples were respectively collected in school children, in which the median urinary iodine was 226.3,282.7 μg/L. One hundred twenty urine samples were respectively collected from the two group of women, resulting a median urinary iodine of 187.2,321.7 μg/L. Eight and 2 salt samples were free of iodine in 120 salt samples collected each region, respectively. The rate of qualified iodized salt was 100%. Conclusions It is not necessary to urgently implement iodine supply in Shouguang City and Licheng District. However, reinforcement of supervise on salt industry and eliminating the hazard from non-iodized and disqualified iodized salt remains in need.

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.