Objective To observe the influence of selenocysteine synthase(SEPSECS) on injury of human umbilical vein endothelial cell line EVC-304 induced by hydrogen peroxide (H2O2). Methods Transfection was conducted to transfect EVC-304 which was maintained in vitro. The cells were divided into four groups: control group, SEPSECS over-expression group, empty vector group and SEPSECS silenced expression group, then Real-time PCR and Western blotting were performed to detected SEPSECS mRNA and protein expression , respectively. Flow cytometry(FCM) was performed to detect cell cycle. Different concentrations of H2O2, which including 0, 200, 400, 600, 800, 1 000 μmol/L, were used to treat EVC-304 . Then malonaldehyde (MDA), superoxide dismutase(SOD) secreted by the cells which were treated with H2O2 for 6 h, were checked by MDA or SOD kit. Results The SEPSECS mRNA expressions of control, SEPSECS silenced expression, empty vector and SEPSECS over-expression groups were 1.03 ± 0.24, 0.43 ± 0.11, 0.98 ± 0.27 and 1.61 ± 0.13, respectively. The protein expressions of control, SEPSECS silenced expression, empty vector and SEPSECS over-expression groups were 1.00 ± 0.26, 0.51 ± 0.10, 1.12 ± 0.38 and 1.51 ± 0.20, respectively. There was a significant difference between control and SEPSECS silenced expression groups (all P<0.05), at the same time , this phenomenon was also observed between empty vector and SEPSECS over-expression groups (all P<0.05). The level of MDA was increased along with the H2O2 concentration. Besides, cell cycle was also tested, however, significant difference was not observed(all P > 0.05). Meanwhile, MDA of SEPSECS silenced expression groups[(15.8 ± 0.5),(19.6 ± 1.5)μmol/L] were significantly higher than control groups[(12.4 ± 0.1),(17.1 ± 0.5)μmol/L, all P < 0.05], on the other hand, MDA of SEPSECS over-expression groups[(10.8 ± 0.4),(14.2 ± 1.1)μmol/L] were lower than empty vector groups [(12.7 ± 0.7),(16.2 ± 1.1)μmol/L, all P < 0.05], when the H2O2 concentration was 800 or 1 000μmol/L. The level of SOD was decreased along with the H2O2 concentration. SOD of SEPSECS silenced expression groups[(7.7 ± 0.4),(2.4 ± 0.3)μmol/L] were lower than control groups[(10.0 ± 1.0),(6.0 ± 0.6)μmol/L, all P < 0.05], on the contrary, SOD of SEPSECS over-expression groups[(11.3 ± 0.6),(12.7 ± 1.6)μmol/L] were higher than empty vector groups[(9.2 ± 0.6),(6.7 ± 0.2)μmol/L, all P < 0.05], when the H2O2 concentration was 800 or 1 000μmol/L. Conclusion Expression of SEPSECS has a significant protective role on damaged EVC-304 which was induced by H2O2.

Objective To measure the erythrocyte dielectric response variation of patients with KaschinBeck disease,to explore the pathogenesis of Kaschin-Beck disease and seek more effective means for evaluating the effectiveness of early diagnosis and control measures against Kaschin-Beck disease.Methods According to the principle of dielectrophoresis and cell dielectric response phenomena,dielectrophoretic pool was manufactured and dielectrophoretic detection and separation system was established.Patients with Kaschin-Beck disease (438) were selected in Kaschin-Beck disease areas of Aba County,Sichuan.At the same time,healthy persons (480) in KaschinBeck disease areas and non-Kaschin-Beck disease areas in Aba County were selected.Anticoagulant blood samples from patients with Kaschin-Beck disease and healthy persons were collected,erythrocytes were separated,cell suspension (10s cells/ml) was prepared and 200 μl cell suspension was added to the dielectrophoretic pool to measure erythrocyte dielectric response rate.Results Erythrocyte dielectric response rates were significantly different between patients living in Kaschin-Beck disease areas with Kaschin-Beck disease [(75.87 ± 5.89)％] and healthy persons living in Kaschin-Beck disease areas and non-Kaschin-Beck disease areas [(92.43 ± 4.45)％,(92.81 ±5.01)％,F =1.843,P ＜ 0.01).Erythrocyte dielectric response rate was significantly reduced in patients with Kaschin-Beck disease compared with that of healthy persons (all P ＜ 0.01);erythrocyte dielectric response rate of adult patients with Kaschin-Beck disease was lower [(69.57 ± 6.87)％] than that of pediatric patients with Kaschin-Beck disease [(82.17 ± 4.91)％,P ＜ 0.01].Erythrocyte dielectric response rates were significantly different in patients with different degrees of Kaschin-Beck disease (F =1.647,P ＜ 0.01).Erythrocyte dielectric response rate of patients with Kaschin-Beck disease was negatively correlated with prevalence of the disease (r =-0.87,P ＜ 0.01).Conclusions Erythrocytes of patients with Kaschin-Beck disease have some degree of pathological damage.The more severe the disease,the more serious the damage of red blood cell.The change of erythrocyte dielectric response properties may be used as an index to judge the prevalence of Kaschin-Beck disease and for early diagnosis.

Objective Through observing the effect of low-dose T-2 toxin on chondrocyte,to study the molecular mechanism of cartilage damage.Methods The primary chondrocytes were isolated from articular cartilage of d 1-2 Wistar neonate rats through enzymatic digestion.Different doses (0.005,0.010,0.100 μg/L) of T-2 toxin were added after 24 h in vitro culture.The survival rate of chondrocytes was detected with Trypan blue staining.Echylosis (matrix metalloproteinase,MMP1) was analyzed by immunohistochemistry.The damage of articular chondrocyte was observed by transmission electron microscope.Results ①Cell morphology of in vitro cultured chondrocyte:the newly isolated chondrocytes were spherical.After 24 hours,the adherent cells gradually began to stretch the triangle or polygon;the nucleus was large and round;the cell was clear and transparent,containing secretory granules.②Cell proliferation:T-2 toxin had a significant inhibitory effect on chondrocyte proliferation,the higher the concentration of T-2 toxin,the significant the inhibitory effects [0.000 μg/L (control) group:3.45 × 108/L,0.005 μg/L T-2 toxin group:3.45 × 108/L,0.010 μg/L T-2 toxin group:2.06 × 108/L,x2 =9.554,P ＜ 0.05].③Immunohistochemical observation:dysplasia,nucleus condensation and membrane rupture were observed in T-2 toxin treated group,brown staining was observed in all groups at varying degrees.The deepest staining was in 0.005 μg/L T-2 toxin group,with the strongest secretion of MMP1;with increasing doses of the toxin,the damage to cartilage cells was severe,MMP1 secretion was less,staining was weak,and the weakest staining was in the 0.100 μg/L T-toxin group.④Under transmission electron microscopy:in control group,cytoplasm was rich in rough endoplasmic reticulum,nuclear membrane and cell membrane were clear;in 0.005 μg/L T-2 toxin group,the cell nucleus showed pyknosis,organelles were decreased in cytoplasm;in 0.100 μg/L T-2 toxin group,the microvilli was dropped out of cartilage surface,nuclear changes were obvious,and mitochondria was myeloid degeneration;rough endoplasmic reticulum was degranulation and expansion into cystiform,chondrocytes were apoptosis occasionally,the cell nucleus showed pyknosis,and the formation of high-density plaque.Conclusion Low dose of T-2 toxin could damage the primary cultured articular chondrocyte in vitro.The results have showed that there are damaged cytostasis,chondrocyte degeneration,necrosis and apoptosis.

Objective To understand the epidemic status quo and influencing factors of Kashin-Beck disease among children in mountain areas of Jilin Province.Methods Two hundred eighty-two severe endemic areas in 18 counties were selected and stratified by random cluster sampling method,and the status quo of KashinBeck disease prevalence was investigated among 7-12 year-old children according to the Diagnostic Criteria of Kashin-Beck Disease (WS/T 207-2010).In the meantime,the annual household income and the proportion of economic crops replanted,grain out-sourced,and returning farmlands to forests and grass were surveyed in the disease affected areas.Results A total of 14 162 children were investigated who had no clinical symptoms.Among them,28 cases were detected positive using X-ray with a detection rate of 1.98‰,while most of the cases were metaphysis positive.The annual household income (≥5 000 Yuan vs.＜ 5 000 Yuan) in the year 2009-2011 had a significant impact on the incidence of Kashin-Beck disease (1.47‰ vs.3.67‰,x2 =6.179,P ＜ 0.05),while the areas of returning farmland to forests and grass which accounted ＞ 1％ had no significant influence on the incidence compared with that ≤ 1％ (3.30‰ vs.1.57‰,x2 =3.876,P ＞ 0.05);the areas of economic crops replanting which accounted ＞ 10％ had no significant influence on the incidence compared with that ≤ 10％ (3.07‰ vs.1.65‰,x2 =2.565,P ＞ 0.05);the proportion of grain out-sourcing which accounted ＞ 50％ had no significant influence on the incidence compared with that ≤50％ (3.07‰ vs.1.65‰,x2 =2.565,P ＞ 0.05).Conehision Up to 2012,the disease among 7-12 year-old children of the mountain areas of Jilin Province have basically met the standard of Kashin-Beck disease elimination and the situation remains stable;furthermore,the household income has a significant impact on the incidence of Kashin-Beck disease.

Objective To observe the expression of neural nicotinic acetylcholine receptor subunit α3 (α3nAChR) and extracellular regulated protein kinases (ERK1/2),c-Jun N-terminal kinase (JNK),p38 kinases of mitogen-activated protein kinase (MAPK) pathway in human neuroblastoma cell line SH-SY5Y overexposed to fluoride,and try to investigate the molecular mechanism of cell damage caused by overexposure of fluoride.Methods The SH-SY5Y cell with low expression of α3nAChR suppressed by silence interference RNA served as α3nAChR silence group;the normal SH-SY5Y cell served as control group,and the effect of silencing of αt3nAChR gene in SHSY5Y was detected by Western blotting and real-time PCR;SH-SY5Y cell was treated with different concentrations of fluoride (0.000,0.005,0.050,0.500,1.000,2.500,5.000 mmol/L),the safe concentration of fluoride in SHSY5Y cell was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay;the SH-SY5Y cell of control group and α3nAChR silence group were treated with 4.000 mmol/L fluoride for 0,4,8,12,24,36,48 h according to the results of MTT assay;the expression of ERK1/2,JNK,p38 kinases of MAPK pathway in SH-SY5Y at protein levels was measured by Western blotting.Results The expression of α3nAChR mRNA (0.04 ± 0.03) and protein (12.0 ± 2.5) in α3nAChR silence group was decreased significantly compared with those of control group (1.00 ± 0.11,100.0 ± 11.3,t =24.58,28.80,all P ＜ 0.05).The viability of SH-SY5Y cell treated with 5.000 mmol/L fluoride (0.53 ± 0.15) was decreased significantly compared with that of SH-SY5Y cell treated with 0.000 mmol/L fluoride (1.05 ± 0.05,P ＜ 0.05).The increased expression of phospho-ERK1/2 was found in α3nAChR silence group and control group incubated with fluoride with time prolonged,and the expression of phospho-ERK1/2 increased significantly at time points 24,36 and 48 h (188.33 ± 7.33,200.00 ± 10.01,213.33 ± 11.55;125.33 ± 5.69,136.00 ± 4.52,155.33 ± 6.51) compared to 0 h in the same groups (100.00 ± 0.00,100.00 ± 0.00,all P ＜ 0.05),and the expression of phospho-ERK1/2 was higher significantly in α3nAChR silence group than those of control group (t =9.26,7.63,5.72,all P ＜ 0.05);no change of expression of total-ERK1/2 in the two groups was found with the passage of time.The gradually increased expression of phospho-JNK was found in α3nAChR silence group and control group,among which,the expression of phospho-JNK in o3nAChR silence group at time points 12,24,36 and 48 h (154.00 ± 6.25,149.00 ± 5.57,156.00 ± 6.08,141.67 ± 2.52) and in control group at 8,12,24,36,48 h (133.33 ± 10.69,173.00 ± 4.00,175.00 ± 11.79,200.67 ± 11.93,200.33 ± 18.58) was compared to those at 0 h in the same groups (100.00 ± 0.00,100.00 ± 0.00),and the difference was significant (all P ＜ 0.05);the higher expression of phospho-JNK was found in α3nAChR silence group other than control group at 8,12,24,36,48 h (t =-4.28,-5.02,-2.89,-8.33,-6.18,all P ＜ 0.05);no change of expression of total-JNK was found in the two groups (P ＞ 0.05).The increased expression of phospho-p38 was detected in control group at time points 24,36 and 48 h (120.33 ± 4.51,122.00 ± 7.55,119.67 ± 7.57) compared to 0 h in the same groups (100.00 ± 0.00,all P ＜ 0.05),and the expression of phospho-p38 was significantly higher than that in α3nAChR silence group at the same time points (93.33 ± 9.61,94.00 ± 5.01,98.33 ± 5.69,t =-4.01,-6.73,-5.59,all P ＜ 0.05);no change of expression of total-p38 was found in the two types of SH-SY5Y cells treated with fluoride (P ＞ 0.05).Conclusion When SH-SY5Y cells are exposed to fluoride;activation of ERK1/2 may be not depend on α3nAChR;α3nAChR may have protected the cell from apoptotic injury caused by activation of JNK pathway,and the activation of p38 may be depend on nAChRα3.

Objective To study the real releasing of fluorine (F) and its effects on environmen in coalburning-borne fluorosis areas.Methods The coal,clay,briquette,cinder,fresh corn and roasted corn samples from 24 households were collected systematically in Zhaotong of City,Yunnan Province,and the F concentration in all samples was measured using the combustion hydrolysis/fluoride-ion selective electrode method,and the released amount and rate of F in briquette was calculated,and the relation between the released amount of F from briquette and the increase of F in roasted corn was analyzed.Results The F concentration of clay ranged from 266.4 to 2 536.7 mg/kg with a median of 1 087.1 mg/kg;the F concentration of briquette ranged from 115.9 to 635.0 mg/kg with a median of 308.8 mg/kg.The released amount of F in briquette ranged from 63.5 to 432.2 mg/kg with a median of 248.2 mg/kg.The releasing rate of F in briquette ranged from 30.3％ to 91.5％ with a average of 73.8％.The F concentration in fresh corn ranged from 0.43 to 2.00 mg/kg with a median of 1.31 mg/kg;the F concentration in roasted corn ranged from 2.21 to 44.88 mg/kg with a median of 10.49 mg/kg.The increased amount of roasted corn ranged 0.90 to 44.46 mg/kg.There was a significant correlation between the emission amounts of F from briquette and the F concentration in binder clay (r =0.714,P ＜ 0.01),but no correlation between the released amount of F from briquette and the increasing of F in roasted corn (r =-0.155,P ＞ 0.05).Conclusion The clay is an important source of F pollution,and the F increase in roasted corn is not only determined by the released amount of F in briquette,but also by several factors.

Objective To establish experimental autoimmune thyroiditis (EAT) rat model induced by bovine thyroglobulin (bTg) and to observe the effect of iodine on IP-10 in rat serum and IP-10 mRNA expression in rat thyroid tissue.Methods One hundred and thirty-five four-week old female Lewis rats were divided into normal control (NC,20 rats) group;TG group,25 rats;H Ⅰ group,20 rats;H Ⅰ + TG group,25 rats;H Ⅱ group,20 rats and H Ⅱ + TG group,25 rats according to a random number table.The water iodine concentration was 25.7 μg/L given to rats of HⅠ and H I + TG groups,and 423.3 μg/L of H Ⅱ and H Ⅱ + TG groups.Rats of NC and TG groups drank distilled water.Rats of TG,H Ⅰ + TG,H Ⅱ + TG groups were immunized with 0.1 ml bTg (8 g/L) in IFA.All rats were killed at the end of 15 weeks.Urinary iodine was determined by As3+-Ce4+ catalytic spectrophotometry (WS/T 107-2006).Pathological changes in thyroid tissue were observed by light microscope.Serum IP-10 was determined by enzyme-linked immunosorbent assay (ELISA),and IP-10 mRNA expression in thyroid was detected by real-time PCR.Results The differences of urinary iodine between groups were statistically significant (x2 =106.4,P ＜ 0.05).Compared with NC group (456.45 μg/L) urinary iodine in other groups increased significantly (TG,H Ⅰ,H Ⅰ + TG,HⅡ,HⅡ + TG:800.08,18 633.20,13 869.08,87 889.97,61 661.51 μg/L,all P ＜ 0.05).The pathological changes of rats in each group aggravated following increased iodine intake,NC group had normal thyroid structure;thyroid of TG group had a small amount of lymphocytes;thyroid of H Ⅰ group showed irregular follicular,part of the follicular was destroyed;a large number of lymphocytes infiltrated between follicular of H Ⅱ group;in H Ⅱ + TG group,the follicular was destroyed severely,diffuse inflammatory cell infiltration.The difference of serum IP-10 between groups no were statistically significant (F =1.462,P ＞ 0.05).The expression of IP-10 mRNA of H Ⅰ + TG (2.80 ± 1.73) rats thyroid was higher than that in NC (1.65 ± 1.62) and H Ⅰ (1.07 ± 1.00) groups,H Ⅱ (0.64 ± 0.64),H Ⅱ + TG (0.80 ± 0.49) were lower than H Ⅰ + TG group (all P ＜ 0.05).Conclusions Excessive iodine intake could have increased inflammatory cells in EAT rat and rats have showed more serious pathologic changes.These phenomena may be ralated with changed expression of IP-10 mRNA in EAT rat thyroid.

Objective To sequence an atypical Brucella strain 16S rDNA,and to evaluate the feasibility of 16S rDNA sequencing method for identification of Brucella.Methods Preliminary identification of atypical strains was carried out with conventional method.Strain DNA was extracted,and 16S rDNA complete sequence was bidirectional sequenced,and Blast in NCBI and DNAMAN software were used for comparison of the sequence identities of the 16S rDNA.Moreover,16S rDNA complete sequence of the stains those were known to cross-react serologically with Brucella was downloaded from GenBank,MEGA 6.0 was used to construct the phylogenetic tree.Results The conventional identification results revealed that it was an atypical Brucella,the gene similarity between the sequences of the test strain 16S rDNA and Brucella was 99％,between 16S rDNA sequence of Brucella abortus 544A and Brucella melitensis 16M was 96.99％.Phylogenetic tree revealed that the test stain was a Brucella,and closely related to Brucella abortus 544A and Brucella melitensis 16M.Conclusion The test strain is an atypical Brucella,and 16S rDNA sequencing analysis is a simple,rapid,and accurate identification method for atypical Brucella.

Objective To study the relationship between land use/cover structure and changes and risk of plague prevalence.Methods Chifeng,Aohan,Alukeerqin and Wengniute were selected as typical areas of Spermophilus Dauricus plague foci in Inner Mongolia,density of Spermophilus Dauricus,plague epidemic and land use data in 1986,1995,2000 were collected,and risk of plague prevalence was represented by the density of Spermophilus Dauricus and epizooty of Spermophilus Dauricus.The percentage of land type structure,change degree of integrated land cover,division index of grassland for plague suitable habitat were calculated with ArcGIS 9.3.The relationship between the land use,cover change and the plague epidemic were analyzed by the methods of comparative.Results From 1982 to 2000,the density of Spermophilus Dauricus was lower than 0.4/hm2 and no epizooty of Spermophilus Dauricus was observed both in Aohan Banner and Chifeng City,which indicated low risk of plague prevalence.Contrarily,from 1982 to 2000,the density of Spermophilus Dauricus was higher than 1.0/hm2,and epizooty of Spermophilus Dauricus was prevalent in Alukerqin Banner,which implied high risk of plague prevalence.In Wenguiute Banner,the density of Spermophilus Dauricus was lower than 0.3/hm2 from 1982 to 1990,after then showed a rising trend,reached higher than 1.0/hm2 in 1991,1995-1997,and epizooty of Spermophilus Dauricus was also monitored,which denoted high risk of plague prevalence;In 1986,1995 and 2000,the proportions of grassland of Wengniute Banner,Alukerqinqi Banner (51.36％,49.14％,48.28％;56.97％,57.48％,57.44％) were much higher than Chifeng and Aohan Banner (35.99％,33.24％,32.95％;38.94％,34.46％,31.31％),but those of arable land (16.81％,18.78％,20.22％;13.28％,14.92％,15.15％) were lower than Chifeng and Aohan Banner (39.73％,42.25％,42.67％;41.21％,43.99％,49.53％).The highest of comprehensively dynamic degree of land cover changes appeared in Aohan Banner,and it was 0.53 and 1.11 in 1986 vs.1995 and 1995 vs.2000,respectively,and the lowest in Alukeerqin Banner,it was 0.22 and 0.05 during those periods.The grassland division index in Alukeerqin Banner was the lowest,which was 0.29 in 1986,0.28 in 1995,and 0.29 in 2000.The grassland division index was higher and showed a upward trends in Aohan Banner,which was 0.57 in 1986,0.69 in 1995 and 0.71 in 2000.Conclusion There is a certain relation between risk of plague prevalence and land use/cover structure and changes.

Objective To know the state of endemic fluorosis,running state of water improvement project in Jiangsu Province,and to provide a scientific basis for adjusting control measures against the disease.Methods Nine key counties were selected from the endemic fluorosis areas as survey spots in Jiangsu in 2012.Three villages in each spot were selected as survey villages.To known the progress and running conditions of water facilities,water samples were collected to determine the fluorosis level.All children aged 8 to 12 were tested for dental fluorosis at the survey villages.The fluorine content in water was determined with F-selective electrode (WS/T 106-1999).Dental fluorosis was diagnosed byased on the Dean method.Results Twenty-seven villages were investigated,the number of villages that had water quality improved was 26,the rate was 96.30％ (26/27);the number of villages that did not have water quality improved was 1,the rate was 3.70％ (1/27).Among the 26 facilities,21 improved-water facilities ran normally,the rate was 80.77％ (21/26);2 facilities ran with intermittently water supply,the rate was 8.70％ (2/26);3 facilities were scraped,the rate was 11.54％ (3/26).In the villages with normal improved-water facilities and qualified water fluorosis,the total rate of dental fluorosis of children aged 8 to 12 was 22.74％ (380/1 761),the index of dental fluorosis was 0.45.In the villages with abnormal improved-water facilities and excessive water fluorosis,the total rate of dental fluorosis of children aged 8 to 12 was 66.51％ (143/215),the index of dental fluorosis was 1.45.The total rate of dental fluorosis of children aged 8 to 12 was 42.86％ (12/28) in the villages which had not changed water source,the index of dental fluorosis was 0.86.Conclusion Improvement of water decreasing fluorine project in Jiangsu Province has a certain effect on children's dental fluorosis,but some of the water fluorosis projects are scrapped,and excess fluorine water should be taken seriously.