In 2012,General Office of the State Council forwarded the'12th Five-Year Plan'on National Prevention and Control of Endemic Diseases,which was jointly developed by the Former Ministry of Health,Development and Reform Commission,and Ministry of Finance.During the 12th Five-Year Plan period,the national prevention and control of endemic diseases had made steady progress.This article mainly summarized the progress made in prevention and control of endemic diseases in China during the 12th Five-Year Plan period.The main problems encountered in prevention and control of endemic diseases in China were discussed.The prevention and key research tasks of the 13th Five-Year Plan on national prevention and control of endemic diseases were introduced.In the 13th Five-Year Plan,key issues in prevention and control of endemic diseases will be the existing main problems,to carry out targeted prevention and control,and to develop scientific and accurate prevention and control plan according to law.

Objective To study the influence of parental generation coal-burning-borne fluorosis on tooth development of their offspring.Methods High fluoride air model was established on the basis of burning coal habit of the epidemic areas.Fluoride feed was made of coal drying corn from the epidemic areas.Totally 48 SD rats were randomly divided into 4 groups with male to female ratio of 1:1 by random number table method.Rats in high,middle and low fluoride groups were put in the high fluoride air room and feed food with 40,25 and 10 mg/kg fluorine,and the control group was put in normal air room and feed normal food.After 8 weeks,rats were mating and parturition.Tooth eruption time of offspring rat was observed;and dental fluorosis incidence,the tooth length and fluorine content were observed at 21 d.Results In high and middle fluoride groups [(6.83 ± 0.94),(6.25 ± 1.06) d],tooth eruption time of offspring rat was later than that of control group [(5.34 ± 0.89) d,all P ＜ 0.01].At 21 d,dental fluorosis was observed in the lower incisors of the high and middle fluorine groups;compared with control group [(5.21 ± 0.19) mm,(223.00 ± 14.08) μg/kg],the tooth length was decreased [(4.83 ± 0.22),(4.96 ± 0.25) mm,P ＜ 0.01or ＜ 0.05],and tooth fluoride content was increased [(362.64 ± 20.35),(289.79 ± 19.18) μg/kg,all P ＜ 0.01].Dental fluorosis incidence of offspring rats was positively correlated with the fluorine dose (r =0.704,P ＜ 0.01).Conclusion Parental generation rats’ intaking excessive fluoride can affect offspring rats tooth development and dental fluorosis,which is related to the fluorine dose.

Objective To observe the influences of NaAsO2 on H3K36me3 modifications,mRNA transcription of O6-methylguanine-DNA methyltransferase gene (MGMT) in HaCaT cells,and to explore the relationship between the transcription of MGMT gene regulated by H3K36me3 and DNA damage induced by arsenic,in order to provide new ideas and scientific basis for prevention and intervention of arsenism.Methods HaCaT cells were treated with 1.25,2.50,5.00 and 10.00 μmol/L NaAsO2 for 24 h,and were also treated with 10.00 μmol/L NaAsO2 for 6,12 and 24 h.HaCaT cells that treated with 0.00 pmol/L NaAsO2 and 0 h were used as blank control group.The degree of DNA damage in peripheral blood cells was detected by single cell gel electrophoresis.The level of H3K36me3 modifications was detected using Western blotting.Quantitative real-time polymerase chain reaction was used to detect the mRNA levels of MGMT gene.Quantitative chromatin immuno-precipitation was used to detect the level of H3K36me3 modifications in the coding regions (ChIP1 and ChIP2) of MGMT gene.Results ①Among the groups of HaCaT cells treated with 2.50,5.00 and 10.00 μmol/L NaAsO2,the levels of tail DNA％ (11.83 ± 1.15,16.85 ± 2.52,24.23 ± 2.75) and olive tail moment (OTM,10.90 ± 1.13,16.19 ± 2.26,23.83 ± 2.79)were significantly increased compared with those of the control group (0.00 μmol/L,2.40 ± 0.51,2.26 ± 0.40,all P ＜ 0.05).After treated with 10.00 μmoFL NaAsO2 for 12 and 24 h,compared with the control group (0 h,3.66 ± 1.02,3.38 ± 1.00),the degrees of tail DNA％ (15.51 ± 1.92,24.18 ± 2.42) and OTM (13.58 ± 2.04,23.14 ± 2.11)were significantly increased (all P ＜ 0.05).②Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the levels of H3K36me3 modifications (60.59 ± 9.75,57.82 ± 11.28,39.45 ± 7.09) were lower at the dosages of 2.50,5.00 and 10.00 μmol/L NaAsO2 (all P ＜ 0.05).Compared with the control group (0 h,100.00 ± 0.00),the levels of H3K36me3 modifications (48.47 ± 9.67,47.75 ± 6.98) were lower after treated with 10.00 μ mol/L NaAsO2 for 12 and 24 h (all P ＜ 0.05).③The levels of H3K36me3 modifications in HaCaT cells exposed to different doses of NaAsO2 were negatively associated with the tail DNA％ and OTM (r =-0.897,-0.903,all P ＜ 0.05).④Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the mRNA levels of MGMT gene were lower at the dosages of 2.50,5.00 and 10.00 pmol/L NaAsO2 (78.20 ± 3.50,61.40 ± 2.60,49.15 ± 4.70,all P ＜ 0.05).⑤There was no observed H3K36me3 enrichmem regularity in the gene encoding ChIP1 and ChIP2 regions of MGMT gene in all doses of NaAsO2 groups (all P ＞ 0.05).Conclusions H3K36me3 may be involved in the regulation of arsenicinduced DNA damage in HaCaT cell.Amenic could inhibit the mRNA transcription of MGMT gene in HaCaT cells,but the transcription of MGMT gene regulate by H3K36me3 is not closely related to DNA damage induced by arsenic.

In recent years,the influence of endemic arsenic poisoning on immune system has caught people's attention.Changes in histone modification patterns may be one of the mechanisms of arsenic regulated gene expression,arsenic plays an important role in differentiation,proliferation and function of immune cells.Arsenic has a certain effect on the differentiation and function of T cells,and it is rarely studied whether the process is regulated by histone methylation.In this paper,the study of histone methylation in arsenic induced T cellular immune damage is reviewed.

Objective To investigate the prevalence of adult skeletal fluorosis in drinking-water-borne endemic fluorosis areas in Liaoning Province and to observe the effects of preventive measures.Methods Three water-changed villages were selected from six drinking-water-borne endemic fluorosis counties,Faku County,Liaoyang County,Haicheng City,Linghai City,Fu Mongolia County,and Jianping County.Totally 18 diseased villages were selected as the investigation sites.The operating condition of the improvement projects was investigated and the fluoride level of drinking water was tested.People over the age of 25 and living in the local for more than 5 years in each survey site were selected and divided into five groups according to age groups of 25-,35-,45-,55-,and ≥65 years old.Ten people in each age group were selected,half male and half female,to examine skeletal fluorosis by X-ray.Clinical symptoms and bone change,including sclerotin,bones,and joints changes were also examined.Results The water fluorine values of two villages were 1.32 and 1.69 mg/L,more than the national standard (1.2 mg/L),while other 16 villages were between 0.5-1.0 mg/L.X-ray images were analyzed for diagnosis of skeletal fluorosis in 905 adults,and 46 cases were diagnosed as skeletal fluorosis in the 18 monitored villages.Xray detection rate was 5.08％,of which the X-ray detection rate of adult fluorosis in water fluoride qualified monitoring villages was 4.11％ (33/803).The detection rate of adult skeletal fluorosis was 12.75％ (13/102) in water fluoride unqualified monitoring villages.There was a significant difference of the total detection rates between the two groups of monitored villages (x2 =13.99,P ＜ 0.05).Skeletal fluorosis was mainly happened in the age group over the age of 45,account for 89.13％ (41/46).Peri-bone changes were observed in the 46 cases of patients with skeletal fluorosis.Conclusions The condition of skeletal fluorosis has been effectively alleviated after implementing the improvement measures.Prevention and treatment can effectively control the occurrence of fluorosis.

Endemic arsenicosis is a disease with complex systemic damage,but the mechanism and its control strategy are mostly focused on a single organ or system damage caused by arsenic,and the systemic damage of arsenic exposure remains not fully understood.In recent years,considerable progress has been made in the field of immunology,which made an important contribution for the pathogenic mechanism of various complex diseases,and also brings a new dawn for its prevention and treatment.In consequence,a breakthrough progress is expected to make in aspects of the mechanism and its control strategy of arsenic caused systemic damage from the perspective of immunology.

Objective To investigate the differential expression levels and significance of regulatory T cell (Treg) and T helper 17 (Th17) related immunologic factors in peripheral blood of arsenic-exposed rats.Methods Thirty-two Wistar rats were numbered by weight,randomly divided into four groups [control,low (1.25 ml/kg),medium (2.50 ml/kg),and high (5.00 ml/kg)],8 rats per group.Rats in control group were given oral gavage of deionized water,and low,medium and high arsenic exposed groups were given oral gavage doses of 2.00 g/L sodium arsenite (NaAsO2) according to their body weight for 6 days every week.After 4 months,the urine and peripheral blood samples of rats were collected,urinary arsenic (uAs) was detected by inductively coupled plasma-mass spectrometry (ICP-MS),the results were shown in [median (minimum and maximum)],uAs was corrected by urinary creatinine (uCr),the unit was μg/g Cr;enzyme-linked immune-sorbent assay (ELISA) was applied to detect the levels of Treg,Th17,T lymphocytes activation related immunologic factors [interleukin-10 (IL-10),transforming growth factor beta1 (TGF-31),IL-17,IL-6,IL-2],the results were shown in (x) ± s.Results The uAs of the rats was compared between control,low,medium,and high arsenic exposed groups [7.50 (3.16-9.81),72.34 (62.34-106.63),209.15 (154.41-232.20),369.73 (289.50-516.55) μg/g Cr],the differences were statistically significant (F =337.55,P ＜ 0.05).IL-10 [(85.03 ± 7.11),(93.96 ± 8.14),(97.48 ± 6.23),(93.47 ± 4.41) ng/L],TGF-β1 [(72.88 ± 2.96),(81.45 ± 8.15),(86.08 ± 7.55),(90.29 ± 5.35) ng/L],IL-17 [(18.15 ± 3.66),(25.54 ± 5.59),(31.48 ± 5.74),(37.25 ± 7.36) ng/L],IL-6 [(83.68 ± 8.48),(85.14 ± 7.11),(89.78 ± 5.36),(93.48 ± 5.77) ng/L],and IL-2 [(80.65 ± 6.90),(73.86 ± 8.00),(69.93 ± 7.77),(62.06 ± 9.82) ng/L] of the rats were compared between control,low,medium,and high arsenic exposed groups,the differences were statistically significant (F =5.094,11.054,16.249,3.474,5.119,all P ＜ 0.05).There were positive correlations between uAs and TGF-β1,IL-17 concentration (r =0.723,0.605,all P ＜ 0.01),while IL-2 showed a negative correlation (r =-0.484,P ＜ 0.05).Concltsion Arsenic exposure could affect the secretion of Treg and Th17 related immunologic factors,so as to the imbalance of anti-inflammatory and pro-inflammatory,which may play a role in the formation and development of arsenic-related immune injury.

Objective To detect the mRNA and protein expression of downstream quinine nicotinamide adenine dinucleotide (NADH) dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) induced by blood nuclearrelated factor E2 (Nrf2) in the peripheral blood of those exposed to arsenic in the endemic area of coal arsenic poisoning in Guizhou Province,and to discuss its role in the process of occurrence and development of liver injury due to coal arsenic poisoning.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey sites,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group based on physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.Peripheral blood samples were collected from all subjects.And mRNA expression of NQO1 and HO-1 were detected by RT-qPCR.Content of NQQ1 and HO-1 were detected by enzyme linked immunosorbent assay (ELISA).Results (①)Results of mRNA expression of NQ01 and HO-1:the relative expression level of mRNA of NQO1 and HO-1 in peripheral blood went up gradually as the degree of liver injury of population exposed to arsenic increased (F =5.548,10.961,all P ＜ 0.05);thereinto,the relative expression level of mRNA of NQO1 of mild hepatosis group (median:0.918) was higher than that of the control group (0.576,P ＜ 0.05),the relative expression level of mRNA of NQO1 of moderately severe hepatosis group (1.243) was higher than those of control group,non-patient group (0.653),non-apparent hepatosis group (0.636) and mild hepatosis group (all P ＜ 0.05);the relative expression level of mRNA of HO-1 of non-patient group (1.059),non-apparent hepatosis group (1.225),mild hepatosis group (1.553) and moderately severe hepatosis group (1.604) were higher than that of control group (0.767,all P ＜ 0.05);the relative expression level of mRNA of HO-1 of mild hepatosis group was higher than that of non-patient group (P ＜ 0.05);the relative expression level of mRNA of HO-1 of moderately severe hepatosis group was higher than those of non-patient group and non-apparent hepatosis group (all P ＜ 0.05).②)Results of protein expression of NQO-1 and HO-1:the level of protein expression of NQO1 and HO-1 in serum went up gradually as the degree of liver injury of population exposed to arsenic increased (F =19.685,17.725,all P ＜ 0.05).Thereinto,the protein expression of NQO1 and HO-1 of non-apparent hepatosis group,mild hepatosis group and moderately severe hepatosis group [NQO1:(6.272 ± 0.744),(6.336 ± 0.628),(6.714 ± 0.540) U/L;HO-1:(45.150 ± 4.813),(45.283 ± 5.049),(48.610 ± 5.365) U/L] were higher than those of control group and non-patient group [NQO1:(5.550 ± 0.730),(5.750 ± 0.427) U/L;HO-1:(39.856 ± 5.249),(42.375 ± 3.014) U/L,all P ＜ 0.05],the protein expression of NQO1 and HO-1 of moderately severe hepatosis group were higher than those of non-apparent hepatosis group and mild hepatosis group (all P ＜ 0.05).Conclusion The expression of mRNA and protein of NQO1 and HO-1 is closely related to the occurrence and development of liver injury due to arsenic exposure in coal arsenic poisoning areas.

Objective To investigate the effect of sodium arsenite (NaAsO2) on the expression of miRNA-21 (miR-21) mediated by transcription factor ETS-1 in human normal hepatocytes (L-02).Methods Dose-effect study:The L-02 cells were treated with different doses of NaAsO2 [0.0 (control),2.5,5.0,10.0,20.0,40.0 μmol/L] for 24 h.Time-effect study:L-02 cells were exposed to 0 (control) and 20 μmol/L NaAsO2 for 12,24,36 and 48 h (n =6).ETS-1 and miR-21 were treated with ETS-1 shRNA and miR-21 inhibitor,respectively.The cells treated with ETS-1 shRNA (100 nmol/L) were divided into 4 groups:①ETS-1 shRNA NC treatment alone (control group);②ETS-1 shRNA NC combined with NaAsO2 (20 μ,mol/L) treatment group;③ETS-1 shRNA treatment alone group;④Treatment with ETS-1 shRNA and NaAsO2 (20 μmol/L) group.The MiR-21 inhibitor (100 nmol/L) treated cells were also divided into 4 groups:① miR-21 inhibitor NC treatment (control group);② miR-21 inhibitor NC combined with NaAsO2 (20 μmol/L);③miR-21 inhibitor group;④miR-21 inhibitor combined with NaAsO2 (20 μ mol/L) treatment group.The expression of ETS-1 mRNA and miR-21 were detected by quantitative real-time PCR (qRT-PCR);the protein expression of ETS-1 was detected by Western blotting.Results Dose-effect study:The expression of ETS-1 mRNA in the groups of 0.0 (control),2.5,5.0,10.0,20.0 and 40.0 μmol/L was 1.008 ± 0.028,1.552 ± 0.029,1.697 ± 0.050,1.842 ± 0.077,2.233 ± 0.096 and 2.235 ± 0.092;miR-21 expression was 1.025 ± 0.094,1.552 ± 0.072,1.683 ± 0.066,1.915 ± 0.171,2.337 ± 0.195 and 2.592 ± 0.177;the expression of ETS-1 protein was 1.060 ± 0.045,1.267 ± 0.160,1.386 ± 0.087,1.723 ± 0.196,2.208 ± 0.122 and 2.284 ± 0.224,respectively,and the differences were statistically significant (F =47.797,8.959,65.748,all P ＜ 0.05),the NaAsO2 dose groups were significantly higher than those of the control group (all P ＜ 0.05),and there was a dose-effect relationship.Time-effect study:The expression of ETS-1 mRNA in L-02 cells was 1.253 ± 0.175,1.623 ± 0.220,1.771 ± 0.324 and 1.913 ± 0.251,respectively at 12,24,36 and 48 h;the expression of miR-21 was 1.502 ± 0.111,1.716 ± 0.113,1.979 ± 0.186 and 2.452 ± 0.304;the expression of ETS-1 protein was 1.196 ± 0.105,1.502 ± 0.076,1.651 ± 0.074 and 1.839 ± 0.139,respectively,there were significant differences between the groups (F =14.936,39.180,39.441,all P ＜ 0.05).The expression of various time points of exposure to NaAsO2 was significantly higher than those in the control group (1.044 ± 0.115,1.044 ± 0.124,1.108 ± 0.088,1.053 ± 0.061;1.092 ± 0.061,1.096 ± 0.169,1.024 ± 0.111,1.057 ± 0.146;1.020 ± 0.017,1.049 ± 0.121,1.024 ± 0.089,1.031 ± 0.124,all P＜ 0.05),and there was a time-effect relationship.ETS-1 shRNA and miR-21 inhibitor treatment:compared with ETS-1 shRNA NC combined with NaAsO2 (20 μmol/L),ETS-1 shRNA and NaAsO2 (20 μmol/L) could significantly inhibit the expression of ETS-1 (0.912 ± 0.238 vs 1.641 ± 0.225,P ＜ 0.05),and down-regulated the expression of miR-21 (1.313 ± 0.334 vs 2.363 ± 0.252,P ＜ 0.05).There was no significant difference of ETS-1 mRNA expression between miR-21 inhibitor and NaAsO2 (20.μmol/L) group (1.580 ± 0.077 vs 1.576 ± 0.065,P ＞ 0.05) compared with miR-21 inhibitor NC and NaAsO2 (20 μmol/L).Conclusions The expression of ETS-1 and miR-21 in L-02 cells is significantly higher than those in control.The high expression of ETS-1 mediates NaAsO2-induced miR-21 overexpression,which may be an important molecular mechanism of arsenic-induced expression dysregulation of human hepatic miRNAs and liver damage.

Objective To observe the activation of rat hepatic stellate cells (HSC-T6) and the changes of methyltransferase (DNMT)1,DNMT3a mRNA expression with different doses of sodium arsenite stimulation.Methods HSC-T6 cells were exposed to a final concentration of 0 (control),5 (low dose),15 (medium dose) and 25 (high dose) μmol/L sodium arsenite in culture medium for 24,48 and 72 h,cells and cell culture supernatant were harvested.Enzyme-linked immunosorbent assay (ELISA) kit was used to measure fibrosis factors contents of type Ⅰ collagen (COL-1),type 11Ⅲ collagen (COL-3) and alpha smooth muscle actin (α-SMA) and quantitative real-time PCR was used to detect the expression levels of DNMT1 and DNMT3a mRNA.Results Different arsenic exposure time (24,48,72 h) had a significant effect on COL-1,COL-3 and α-SMA contents in HSC-T6 cells (F =249.574,328.493,3 157.436,all P ＜ 0.01);Different arsenic exposure content (low,medium,high dose groups) had a significant effect on COL-1,COL-3 and α-SMA contents in HSC-T6 cells (F =3 946.521,1 006.399,13 025.770,all P ＜ 0.01).After arsenic exposure for 24 and 48 h,the expression levels of DNMT1 mRNA in high dose group (4.33 ± 0.24,2.34 ± 0.43) were higher than those of control group (1.00 ± 0.00,1.00 ± 0.00,all P ＜ 0.05).At the same arsenic exposure levels (low,medium or high dose),the expression level of DNMT1 mRNA was declined with prolongation of sodium arsenite stimulation time (all P ＜ 0.05).After arsenic exposure for 48 and 72 h,the expression levels of DNMT3a mRNA in high dose group (2.23 ± 0.50,5.02 ± 0.23) were higher than those of control group (1.00 ± 0.00,1.00 ± 0.00,all P ＜ 0.05).The expression levels of DNMT3a mRNA in medium and high dose groups at 72 h (3.80 ± 0.14,5.02 ± 0.23) were higher than those of 24 h (3.03 ± 0.12,0.42 ± 0.15,all P ＜ 0.05).Conclusion HSC-T6 cells are obviously activated with pro-fibrotic effect;the expression levels of DNMT1 and DNMT3a mRNA are both up regulated in HSC-T6 cells after being exposed to sodium arsenite.