Objectives To study the prevailing status of coal-burning type endemic fluorosis in Fuyuan County of Yunnan Province, and to provide a scientific basis for fluorosis control and prevention. Methods In 2013, five towns were selected from the villages and towns which the defluoridation stove project was implemented and by the end of 2007 defluoridation stove rate was higher than 95%(including 95%), and two natural villages were selected in each township to carry out the investigation. Dental fluorosis was examined of all children aged 8 to 12 of survey points. At least 20 people were selected in each village. At the same time, 20 copies of instant urine samples were collected from these children of half male and half female, and urinary fluoride was determined. Then five natural villages were sampled from all survey points, and permanent residents were divided into25-,35-,45-,55-and≥65groups according to age. Six people were selected from each group with male and female in half who were conducted examination of adult skeletal fluorosis by X-ray. Dental fluorosis diagnosis was based on theDental Fluorosis Diagnosis(WS/T 208-2011). Urinary fluoride determination was done in accordance withUrine-Determination of Fluoride-Ion Specific Electrode Method(WS/T 30-1996). Skeletal fluorosis diagnosis was carried out according to Diagnostic Criteria for Endemic Skeletal Fluorosis (WS 192-2008). Results A total of 556 children aged 8 to 12 were monitored, the detection rate of dental fluorosis was 41.55%(231/556), the dental fluorosis index was 0.78. The very mild, mild, moderate and severe proportions of children's dental fluorosis were 20.50%(114/556), 17.09%(95/556), 3.60%(20/556) and 0.36%(2/556), respectively, given priority to very mild. The prevalence of dental fluorosis in children was increased with increasing age (χ2=74.27, P<0.05). One hundred and ninety-three copies of child urine samples were tested, the geometric mean of urinary fluoride was 0.62 mg/L, and the urinary fluoride contents were between 0.10-2.93 mg/L. A total of 116 adults were examined, and the detection rate of skeletal fluorosis was 9.48%(11/116). Detected cases of skeletal fluorosis was given priority to mild (9 cases, accounted for 81.81%), more men cases [16.36% (9/55)] than those of women [3.28% (2/61), χ2 = 5.77, P< 0.05]. Conclusion The hazard of coal-burning type fluorosis has been reduced in Fuyuan County , the affect of defluoridation stove project is obvious.

Objective:To investigate the therapeutic effect of conjoint fascia sheath suspension and levator palpebrae muscle shortening on congenital severe blepharoptosis.Methods:From June 2014 to December 2018, 30 cases (40 eyes) of congenital severe ptosis were treated in Ningbo First Hospital and Shaoxing Women and Children＇s Hospital. All patients were corrected by conjoint fascia sheath suspension and levator palpebrae muscle shortening. Six months after operation, the distance between the middle point of upper eyelid margin and corneal reflex point was measured to evaluate the correction effect of blepharoptosis; the improvement of upper eyelid appearance was evaluated by 5-point Likert scale (LS), and the incidence of complications was counted.Results:Thirty patients (40 eyes) were followed up for 6-12 months. Correction effect showed that preoperative marginal reflex distance (MRD) was (0.10±0.05) and postoperative MRD was (3.80±0.55); the difference was statistically significant ( t=0.95, P<0.05); 38 eyes (95%) were corrected and 2 eyes (5%) were undercorrected; appearance of upper eyelid showed that preoperative LS was (0.50±0.05) and postoperative LS (3.80±0.55); the difference was statistically significant ( t=0.98, P<0.05). Only one case was complicated with keratitis caused by improper early nursing after incomplete closure exposure, and the patient was cured by sealing the eyes combined with drug conservative treatment. Conclusions:Combined conjoint fascial sheath suspension and levator palpebrae shortening in the treatment of severe blepharoptosis has significant effect, with the advantages of simple operation, small surgical trauma, low recurrence rate, low incidence of complications, high predictability of surgical results and high satisfactory rate of patients.

Objective:To observe the effect of dictyophora polysaccharide (DIP) on PINK1/Parkin pathway mediated mitophagy induced by sodium arsenite (NaAsO 2) in human hepatocytes (L-02 cells). Methods:The L-02 cells in logarithmic growth phase and in good condition were divided into control group, NaAsO 2 group (10 μmol/L), DIP group (80 μg/ml), DIP + NaAsO 2 group (80 μg/ml DIP + 10 μmol/L NaAsO 2) , N-acetylcysteine (NAC) group (5 mmol/L), and NAC + NaAsO 2 group (5 mmol/L NAC + 10 μmol/L NaAsO 2). Western blotting was used to detect the expression levels of mitophagy related proteins p62, microtubule-associated protein 1 light chain 3 (LC3)Ⅱ/LC3Ⅰ, PINK1, and Parkin. The mitochondrial stucture and autophagosomes were observed by transmission electron microscope, the fluorescent probe method was used to detect the expression level of intracellular reactive oxygen species (ROS). Results:Compared with the control group, the protein expressions of p62, LC3 Ⅱ/LC3 Ⅰ, PINK1, and Parkin in NaAsO 2 group were higher ( P < 0.05); compared with the NaAsO 2 group, the protein expressions of p62, LC3 Ⅱ/LC3 Ⅰ, PINK1 and Parkin were lower in DIP, DIP + NaAsO 2, NAC, and NAC + NaAsO 2 groups ( P <0.05). According to the transmission electron microscope, compared with the control group, the mitochondria of L-02 cells in NaAsO 2 group were significantly damaged and the number of autophagosomes increased. Compared with NaAsO 2 group, the degree of mitochondrial swelling, vacuolar degeneration and the number of autophagosomes decreased in DIP + NaAsO 2 group. Compared with the control group (33 110.00 ± 2 191.28), the intracellular ROS level in NaAsO 2 group was higher (48 000.00 ± 2 395.31, P < 0.05); the level of intracellular ROS in DIP + NaAsO 2 group (38 670.00 ± 2 620.56) was significantly lower than that in NaAsO 2 group( P < 0.05), and there was no significant change compared with the control group ( P > 0.05). Conclusions:NaAsO 2 can induce PINK1/Parkin mediated mitophagy in L-02 cells. DIP can alleviate NaAsO 2 induced mitophagy. DIP may affect PINK1/Parkin mediated mitophagy induced by NaAsO 2 through the regulation of ROS.

Objective To investigate the status of coal-burning type endemic fluorosis in Ludian County of Yunnan Province,to evaluate the prevention and control effect.Methods Multi-stage sampling was adopted in 2016.Five townships from Ludian endemic fluorosis areas and 2 villages of each township were randomly selected.The condition (children's dental fluorosis,urinary fluoride,adult skeletal fluorosis),epidemic factors (use of stoves,fluoride content of main household foods,indoor air fluoride content),health education awareness of endemic fluorosis were investigated in the above mentioned villages.Results The dental fluorosis positive rate of children aged 8-12 was 10.14％ (51/503) while a total of 503 children were investigated,the index of dental fluorosis was 0.16,and adult skeletal fluorosis detection rate was 1.92％ (2/104) while a total of 104 adults were investigated.The geometric mean of urinary fluoride content of children was 0.39 mg/L,ranging from 0.02 to 2.54 mg/L.Totally 98,161,and 151 samples of pepper,rice,and corn were detected.The medians of fluoride content were 0.96,0.42,and 0.43 mg/kg,respectively,ranging from 0.26 to 13.46,0.13 to 2.20,and 0.21 to 2.54 mg/kg,respectively,in which the fluorine contents of pepper 48.98％ (48/98) was higher than 1.0 mg/kg.The correct drying rates of both corn and pepper were 100.00％ (201/201),and the correct preservation rates of corn and pepper was 99.50％ (200/201) and 100.00％ (201/201),respectively.The use rate of improved stove,rate of intact stove,correct use rate of intact stove were 100.00％ (201/201),96.02％ (193/201) and 100.00％ (193/193),respectively.The fluoride contents in the indoor air of the families (the fire period was 0.32 μg/m3 and the fire extinguishing period was 1.22 μg/m3) from the areas of fluorosis which had properly used the improved stoves were lower than that of the national hygienic limit content (the one-hour average concentration limit is 20 μg/m3).The knowledge of prevention and control of coal-burning type fluorosis among fifth-grade primary school students and household heads was investigated in 104 and 201 people,respectively,with the awareness rates of 80.77％ (252/312) and 86.23％ (520/603),respectively.Conclusion By implementing the comprehensive prevention and control measures,which mainly focus on the improvement of the stove and the health education,the coal-burning fluorosis in Ludian County has been effectively controlled,and the prevention and control work has achieved remarkable results.

Objective:To explore the mechanism of apoptosis induced by sodium arsenite (NaAsO 2) in human hepatic cells (L-02) through reactive oxygen species (ROS) accumulation and mitochondrial dysfunction, and provide experimental evidence for the mechanism of arsenic poisoning. Methods:L-02 cells were divided into control group, NaAsO 2 group (10 μmol/L NaAsO 2), N-acetylcysteine (NAC) group (5 mmol/L NAC), and NaAsO 2 + NAC group (10 μmol/L NaAsO 2, 5 mmol/L NAC), and were cultured in vitro for 24 h. The intracellular ROS level, mitochondrial membrane potential depolarization ratio and cell apoptosis rate were measured by dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence probe, JC-1 staining and Annexin V-FITC/PI double staining, respectively; the mRNA and the protein of Caspase 3, cytochrome C (Cyt-C) and cytochrome C oxidaseⅣ (COXⅣ) were detected by real time fluorescence quantitative PCR (qRT-PCR) and Western blotting, respectively. Results:There were statistically significant differences in intracellular ROS levels (3 857 392.33 ± 44 928.39, 4 515 288.00 ± 32 660.64, 3 670 150.67 ± 101 987.69, 4 035 235.67 ± 99 995.30), mitochondrial membrane potential depolarization ratios (2.16 ± 0.54, 7.95 ± 0.52, 2.70 ± 0.29, 1.01 ± 0.23) and total apoptosis rates (1.45 ± 0.03, 4.27 ± 0.17, 1.87 ± 0.12, 2.52 ± 0.35) between groups ( F = 62.62, 159.81, 112.70, P < 0.05). There were statistically significant differences in Caspase 3, Cyt-C, COXⅣ mRNA expression levels ( F = 9.20, 7.33, 14.87, P < 0.05) and in cleaved-Caspase 3, Cyt-C, COXⅣ protein expression levels( F = 31.42, 8.01, 83.30, P < 0.05) between groups. Compared with the control group, the intracellular ROS level, mitochondrial membrane potential depolarization ratio and total apoptosis rate were significantly increased ( P < 0.05); Caspase3, Cyt-C mRNA and protein expression levels were significantly increased ( P < 0.05), and COXⅣ mRNA and cleaved-Caspase 3, Cyt-C protein expression levels were significantly decreased ( P < 0.05) in NaAsO 2 group. Compared with the NaAsO 2 group, the intracellular ROS level, mitochondrial membrane potential depolarization ratio and total apoptosis rate of NaAsO 2 + NAC group were significantly decreased ( P < 0.05); the Caspase3, Cyt-C mRNA and cleaved-Caspase 3, Cyt-C protein expression levels were significantly decreased ( P < 0.05), the COX Ⅳ mRNA and protein expression levels were significantly increased ( P < 0.05). Conclusions:NaAsO 2 stimulates L-02 cells to produce excessive ROS, which induces mitochondrial depolarization and further triggers mitochondrial damage, resulting in increased release of Cyt-C and activation of the mitochondrial apoptosis pathway that Caspase 3 protein induces apoptosis in L-02 cells, which may be one of the main mechanisms of arsenic-induced liver injury.

Objective:To explore the role of nuclear factor-E2-related factor 2 (Nrf2) signaling pathway in oxidative damage caused by sodium arsenite (NaAsO 2) in human normal liver cells (L-02), and to provide experimental basis for the study of oxidative damage mechanism of liver damage caused by arsenic. Methods:L-02 cells were cultured in vitro and treated with 0 (control), 25, 50, 75, 100, 125, and 150 μmol/L NaAsO 2, respectively, for 24 h. The half-inhibitory concentration (IC 50) was calculated according to the cell survival rate by CCK8, and L-02 cells were treated with 0, 1/8, 1/4 and 1/2 dose of IC 50 of NaAsO 2, respectively, for grouping experiments. Protein expressions of Nrf2, heme oxygenase-1 (HO-1), NADH quinone oxidoreductase 1 (NQO1) and glutathione peroxidase 1 (GPx1) in L-02 cells and L-02 nucleus were detected by Western blotting. Results:The result of CCK8 showed that the survival rates of L-02 cells in 25, 50, 75, 100, 125, 150 μmol/L NaAsO 2 groups were [(69.53 ± 0.06)%, (41.33 ± 0.08)%, (23.65 ± 0.04)%, (26.51 ± 0.04)%, (31.63 ± 0.01)%, (26.24 ± 0.02)%], which were significantly lower than that of the control group[(100 ± 0.00)%]. The differences were statistically significant ( P < 0.05). The IC 50 calculated by cell survival was 40 μmol/L, and the NaAsO 2 doses used in the experiment were 0 (control), 5, 10, and 20 μmol/L. Western blotting results showed that, compared with the control group, the protein expression levels of Nrf2, HO-1 in L-02 and HO-1 in the L-02 cells nucleus in the 5, 10 and 20 μmol/L NaAsO 2 groups were significantly higher ( P < 0.05). Compared with the control group, the expression levels of GPx1 protein in L-02 cells of 10 and 20 μmol/L NaAsO 2 groups were decreased ( P < 0.05). Compared with the control group, the expression levels of Nrf2 protein in L-02 nucleus in 10 and 20 μmol/L NaAsO 2 groups were significantly increased ( P < 0.05); the expression level of NQO1 protein in L-02 nucleus in 5 μmol/L NaAsO 2 group was significantly increased ( P < 0.05). Conclusion:NaAsO 2 has an effect on the expression of Nrf2 signaling pathway related factors in L-02 cells, and the mechanism of oxidative damage caused by NaAsO 2 in L-02 cells may be related to Nrf2 signaling pathway.

Objective:To investigate the effects of sodium arsenite (NaAsO 2) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c), peroxisome proliferator activated receptor α (PPARα) and fatty acid synthase (FAS) in human liver cells (L-02 cells). Methods:L-02 cells were cultured in vitro, and exposed to NaAsO 2 at 0 (control), 2, 4, 8, 16, 32, 64 and 128 μmol/L for 24 h, respectively, and the cell survival rate was determined by CCK-8 method. And a fitting curve was made to calculate the half inhibitory concentration (IC 50), subsequent experiments were carried out with 0, 1/8, 1/4 and 1/2 of IC 50 as arsenic exposure doses. Glycerol phosphate oxidase-catalase (GPO-PAP) method was used to detect the content of triglyceride (TG) in cells; the mRNA expression levels of SREBP-1c, PPARα and FAS were detected by Real-time PCR; and the protein expression levels of SREBP-1c and PPARα were detected by Western blotting. Results:The cell survival rates of 8, 16, 32, 64 and 128 μmol/L NaAsO 2 groups [(92.000 ± 1.414)%, (91.000 ± 0.000)%, (76.500 ± 0.707)%, (53.000 ± 1.412)%, (47.000 ± 1.412)%] were significantly lower than that of the control group [(100.000 ± 0.000)%, P < 0.01]. The IC 50 was 64 μmol/L, and subsequent experiments were conducted with 0 (control), 8, 16 and 32 μmol/L NaAsO 2, respectively. Compared with the control group [(1.000 ± 0.000) mmol/g prot], TG contents of 8, 16 and 32 μmol/L NaAsO 2 groups [(0.691 ± 0.064), (0.474 ± 0.162), (0.184 ± 0.045) mmol/g prot] were significant decreased ( P < 0.01). Compared with the control group, the mRNA expression levels of SREBP-1c, PPARα, FAS, and the protein expression levels of SREBP-1c and PPARα in NaAsO 2 groups were significantly decreased ( P < 0.01 or < 0.05). Correlation analysis showed that NaAsO 2 content was negatively correlated with TG content, SREBP-1c and PPARα protein expression levels ( r =-0.954,- 0.875,-0.965, P < 0.01). Conclusion:NaAsO 2 can reduce the TG content and the expression of lipid metabolism related genes SREBP-1c, PPARα and FAS in L-02 cells, suggesting that arsenic-induced liver injury can cause lipid metabolism disorders.

Objective:To accurately grasp the current status of water iodine distribution in environmental of Yunnan Province, and provide a basis for scientific iodine supplementation.Methods:In 2017, according to the "National Drinking Water Iodine Content Survey Program", a survey of water iodine content was conducted in townships. In 2018, a search survey was carried out for administrative villages with water samples of iodine content > 40 μg/L in drinking water found in previous surveys. In centralized water supply villages, 2 samples of end water were collected. In decentralized or partially decentralized water supply villages, each village was divided into 5 parts: east, south, west, north and center. In villages with more than 50 water sources, 10% of the water sources were selected from each part; villages with less than 50 water sources, one water source was selected from each part; villages with less than 5 water sources, all the water sources were selected to determine the water iodine content. At the same time, the mode of water supply and demographic data were investigated.Results:At the township level water iodine survey, a total of 24 589 water samples were collected from 1 389 townships in 129 counties of 16 prefectures (cities). The median water iodine content of township was 1.3 μg/L, ranging 0.0 - 9.3 μg/L. The population covered by the survey accounted for 53.94% (25 013 804/46 371 207) of the total population of the province. According to the survey results of high iodine villages, 93 administrative villages in 68 townships of 37 counties in 12 prefectures (cities) were surveyed, covering 40 940 households and 208 880 people. A total of 761 samples of drinking water were tested, and the median water iodine content of administrative villages was 1.3 μg/L (ranging 0.0 - 196.8 μg/L). There were 2 administrative villages with iodine higher than 100 μg/L, which were Xiaobona Administrative Village (145.8 μg/L) in Liuchang Township, Xiangyun County, and Dasongshu Administrative Village (196.8 μg/L) in Yangchang Township, Xuanwei City.Conclusions:The iodine content of water in Yunnan Province is generally low, which belongs to the area of environmental iodine deficiency. For the high iodine villages, further investigation should be carried out to clarify the prevention and control measures.

Objective To analyze the iodine nutritional status in the key populations before and after the adjustment of iodized salt in Yunnan Province,and to provide scientific basis for adjusting the strategy of prevention and treatment timely.Methods The probability proportional to size sampling method was employed in the investigation.In the pre-adjustment period (2011) and the post-adjustment period (2014),the changes in the residents' iodized salt,the urinary iodine and goiter prevalence of children aged 8-10,the urinary iodine of pregnant women and lactating women were analyzed.Results Before and after adjusting the salt iodine concentration,the coverage rate of iodized salt was 99.1％ (1 196/1 207) and 99.2％ (1 532/1 545),respectively,the mean of salt iodine after the adjustment (23.6 mg/kg) was significantly lower than that of before (30.1 mg/kg,F =17.287,P ＜ 0.01).There was significant difference in the iodine nutritional status of children from 271.4 to 180.9 μg/L (Z=-12.883,P ＜ 0.01).The difference of iodine nutritional status in pregnant women between pre-adjustment (217.3 μg/L) and postadjustment (143.7 μg/L) was also significant (Z =9.997,P ＜ 0.01).The thyroid goiter rate of children had inceased from 1.7％ (21/1 207) to 2.1％ (33/1 549),the difference was not statistically significant between the groups (x2=0.539,P ＞ 0.05).Conclusions After adjusting the salt iodine concentration in Yunnan Province,iodine nutrition of children has decreased from more than adequate level to adequate level,thyroid goiter rate of children has remained at the low level,and iodine nutrition of pregnant women is sightly lower than adequate level,but iodine deficiency of pregnant women is at a low risk because of the good iodized salt coverage.The new standard of iodized salt is appropriate,and it is more favorable to health in Yunnan Province.

Objective To understand the situation of iodine deficiency disorders and iodine nutrition of children in Yunnan Province after the implementation of new standard of salt iodine,and provide the basic data for prevention and control of iodine deficiency disorders.Methods Using multistage sampling method,Yunnan provincial evaluations for iodine deficiency disorders elimination were carried out.According to the comparative analysis of 2010 (pre-adjustment) and 2015 (post-adjustment) evaluation results,the changes of the median of salt iodine,the coverage rate of iodized salt,the qualified rate of iodized salt,the rate of qualified iodized salt consumption,goiter rate of school children aged 8-10 and the median of urinary iodine were evaluated.Salt iodine was tested using the "Salt Industry General Test Method for Determination of Iodine" (GB/T 13025.7-1999);thyroid volume examination of children using B-scan ultrasonography,thyroid volume calculation and enlargement judgment using "Diagnostic Criteria for Endemic Goiter" (WS 276-2007);and urinary iodine detection using "Urinary Iodine Arsenic Cerium Catalytic Spectrophotometry" (WS/T 107-2006).Results The medians of salt iodine in 2010 and 2015 were 30.6 and 22.4 mg/kg,respectively,the difference was statistically significant (U =317 503.50,P ＜ 0.01).The coverage rates of iodized salt in 2010 and 2015 were 99.6％ (1 681/1 688) and 98.9％ (2 592/2 622),the qualified rates of iodized salt were 98.6％ (1 657/1 681) and 89.7％ (2 325/2 592),and the rates of qualified iodized salt consumption were 98.2％ (1 657/1 688) and 88.7％ (2 325/2 622),and the differences were statistically significant (x2 =6.42,125.11,130.18,P ＜ 0.01);the rates of children goiter in 2010 and 2015 were 0.8％ (25/3 272) and 1.4％(44/3 245),respectively;the medians of urinary iodine of children were 289.0 and 157.3 μg/L,respectively,the difference was statistically significant (U =1 121 669.50,P ＜ 0.01).Conclusion After decreasing of salt iodine level starting from 2012 in Yunnan Province,the children iodine nutrition condition is in a more proper level.