To investigate the effect of GFP fused to C terminal of integrin alpha(IIb) on the biosynthesis and expression of alpha(IIb) beta(3) compound, the alpha(IIb) GFP expression plamid, named palpha(IIb) GFP, the cDNA of alpha(IIb) was constructed from p3.1-2b and fused to pEGFP-N1 in frame. When the sequence of palpha(IIb) GFP was confirmed by sequencing it was transferred to Chinese Hamster Ovary (CHO) cells with or without p3.1-3a expressing integrin beta(3). Then the expression of alpha(IIb) GFP fusion protein was confirmed by Western blot and then its subcellular localization was determined with laser confocal scanning microscopy. The results showed that the target gene was cloned into recombinant vector by restriction analysis and sequencing. Overexpression of the fusion protein in the transfected CHO cells was identified with Western blot. Subcellular localization analysis confirmed that alpha(IIb) GFP was expressed in CHO cells and could be transferred from endoplasmic reticulum to Golgi apparatus. It is concluded that the eukaryotic expression plasmid containing alpha(IIb) GFP fusion gene is successfully constructed. GFP fused to the cytoplasmic tail of integrin alpha(IIb) allows the normal expression of alpha(IIb) beta(3) in CHO cells.
Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Endoplasmic Reticulum ; metabolism ; Golgi Apparatus ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Platelet Glycoprotein GPIIb-IIIa Complex ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Objective To observe the illness change trend of drinking water type endemic arsenic poisoning in Qinghai Province, comprehensively evaluate the effect of prevention and control measures, in order to provide a scientific basis for timely adjustment of the prevention and control measures. Methods From 2010 to 2013, according to the Drinking Water Type Endemic Arsenic Poisoning Monitoring Programs, 3 villages in 2 counties within Qinghai Province were randomly selected as monitoring sites. The arsenic content in drinking water of residents was measured, water improvement projects in all monitoring villages were investigated; at the same time an investigation of arsenic disease in resident population was conducted, and urinary arsenic content was monitored. Arsenic in drinking water and urine was determined by hydride generation atomic fluorescence spectrometry, and arsenic poisoning was diagnosed using Endemic Arsenic Poisoning Diagnostic Criteria (WS/T 211-2001). Results Of the three water improvement projects, two were water arsenic exceeded the standard, and one was intermittently operated. From 2010 to 2013, the arsenic poisoning detec tion rate in Baoning Village was 27.30% (193/707), 31.90%(245/768), 29.35%(221/753) and 28.22%(219/776); in Kecai Village was 32.62%(107/328), 34.83%(124/356), 31.26%(131/419) and 29.35%(118/402);and in Manimotai Village was 56.58%(43/76), 52.81%(47/89), 45.10%(46/102) and 34.69% (34/98), there was no significant difference statistically of the arsenic poisoning detection rates in the three monitored villages in the 4 years (χ2 =3.09, 0.04, 0.92, all P>0.05). From 2010 to 2013, women arsenic poisoning detection rate was 36.45%(203/557), 40.59%(246/606), 36.12%(225/623) and 34.77%(218/627), respectively;men was 25.27%(140/554), 28.01%(170/607), 26.57%(173/651) and 23.57%(153/649), respectively;women arsenic poisoning detection rates were higher than those of men (χ2 = 16.25, 21.32, 13.49, 19.38, all P < 0.05). Arsenic poisoning detection rate of people younger than 60 years old had a tendency to increase with age. In 2012 and 2013, 105 and 93 urine samples were tested, respectively; urinary arsenic geometric mean was 0.113 and 0.149 mg/L. Conclusions Water improvement projects and water quality are not optimistic, and prevalence of arsenic poisoning is still at a higher level. A sound long-term monitoring program should be established as soon as possible, the management and maintenance of water improvement projects should be strengthened, and the monitoring and prevention work should not be neglected.

Huacao Community Health Service Center has established an integrated chronic kidney disease (CKD) management system since 2015,which greatly improved CKD management in the community.From September 2016 to August 2017,4084 residents were screened by the family doctor team,975 were suspected to have CKD (23.87％),among whom 202 (20.72％) were referred to district CKD treatment center,and 197 (97.52％) were confirmed as CKD.The detection rate of suspected CKD among patients with hypertension and diabetes mellitus was the highest,which was much higher than that among outpatients and those with routine health check up.It is suggested that the integrated management of CKD should be widely implemented to cope with the prevalence of CKD,particularly for those with hypertension and diabetes.

Objective To explore the prevalence state of drinking brick-tea type fluorosis in children in Guoluo Prefecture,Qinghai Province.Methods According to the historical prevalence of drinking tea-type fluorosis in Guoluo Prefecture,in 2017,in 5 counties of Guoluo Prefecture,according to the pastoral area,agricultural area,semi-agricultural and semi-pastoral areas,each township (town) was selected,in each township (town) drawed a natural village in a ward as a survey point.At each survey site,drinking water samples of local residents were collected,and 10 households were selected,tea samples were collected,and dental fluorosis was investigated for children aged 8-12 years old.The mean of fluoride was determined by fluoride ion selective electrode method (GB/T 5750.5-2006,GB 19965-2005),and the dental fluorosis was diagnosed based on "Diagnosis of Dental Fluorosis" (WS/T 208-2011).Results The mean of fluoride in 85 water samples was 0.34 mg/L (0.20-0.77 mg/L),the mean of fluoride in 161 brick-tea samples was 579 mg/kg (110-1 278 mg/kg).The mean of daily fluoride intake of each person was 6.78 mg,among them,73.29％ (118/161) drank brick-tea,the detection rate of dental fluorosis of 2 083 children was 22.85％ (476/2 083),and the index of dental fluorosis was 0.42,the prevalence was weak.Conclusion The prevalence of drinking brick-tea type fluorosis of children in Guoluo Prefecture is less serious.

Objective:To investigate the contents of soil metal elements in the plague high incidence area and resting area in the plague natural foci of Qingnan region in Qinghai Province, and to compare and analyze the differences of the contents of soil metal elements between the two areas.Methods:A total of 69 soil samples were collected from 11 sites in the plague high incidence area (Yushu Prefecture) and resting area (Guoluo Prefecture) in Qingnan region, including 39 samples from high incidence area and 30 samples from resting area. The contents of 12 soil metal elements, such as iron (Fe), magnesium (Mg), calcium (Ca), barium (Ba), cobalt (Co), copper (Cu), manganese (Mn), lead (Pb), rubidium (Rb), titanium (Ti), vanadium (V) and zinc (Zn), were measured by X-ray fluorescence spectrometry and SPSS 20.0 software was used for Spearman correlation and other statistical analysis.Results:The contents of Fe and Ca in the soil of the plague natural foci of Qingnan region were relatively high. The contents of 12 soil metal elements in the high incidence area were significantly higher than those in the resting area ( P < 0.01 or < 0.05). There were statistical significant differences in the contents of Mg, Ca, Ba, Mn, Pb and Zn in the soil among the sample plots of the plague high incidence area ( P < 0.01 or < 0.05). There were statistical significant differences in the contents of Fe, Mg, Ca, Rb and Ti in the soil among the sample plots of the plague resting area ( P < 0.05 or < 0.01). Except for Mg and Ca, Fe was positively correlated with other metal elements in the high incidence area and resting area ( P < 0.01 or < 0.05), and Fe was strongly positively correlated with Rb, Ti and V ( r = 0.780 - 0.838). Mg was only positively correlated with Ca in the high incidence area and resting area ( P < 0.01). Ca was negatively correlated with Mn, Pb, Rb, Ti, V and Zn in the high incidence area, and with Rb and Zn in the resting area ( P < 0.01 or < 0.05). The rest of the metal elements were mostly positively correlated in the high incidence area and resting area. Conclusions:The contents of Fe and Ca are rich in the plague natural foci of Qingnan region in Qinghai Province. There are correlations among most soil metal elements in the high incidence area and resting area. The correlation between the contents of soil metal elements in the plague natural foci and the epidemic of plague is worth further exploring.

Objective To dynamic monitor drinking-water-borne fluorosis in Huzhu County and water improvement projects,to know the trend of the disease,and to evaluate the effect of prevention and control measures.Methods Seven townships in Huzhu County including 60 history drinking-water-borne fluorosis villages were selected as investigation sites in 2017,and demographic data were collected.In the villages of the diseased areas where the water had been changed,the operation of the water improvement project was investigated,and 1 tap water sample was collected to determine the fluorine content;for the villages in the diseased areas where the water had not been changed,one water sample was collected from the drinking water source in accordance with the five directions of east,west,south,north and middle to determine the fluorine content.More than 90％ of the children aged 8-12 years old in the surveyed villages were examined and judged for dental fluorosis.According to historical data,1 village with severe illness of children was selected,50 people were selected who were over 25 years old and lived in the villages for more than 5 years.Clinical and X-ray examinations of skeletal fluorosis were performed.Urine samples from adults surveyed were collected for urinary fluoride detection.Results Six of the 60 villages in the 7 townships had been relocated.By 2017,a total of 54 villages in drinking-water-borne fluorosis areas had changed their water.A total of 16 water improvement project were surveyed,among them,there were small water improvement project 6,accounting for 37.50％,large water improvement project 10,accounting for 62.50％;normal operated project 16,accounting for 100.00％;water fluoride content of qualified project 16,the qualified rate was 100.00％,the water fluoride content ranged 0.10-0.66 mg/L,covering a population of about 160 thousand and 846 people.A total of 2 399 children aged 8-12 were examined,117 cases of dental fluorosis were detected,the detection rate of dental fluorosis was 4.88％,dental fluorosis index was 0.13.Among them,89 cases were extremely mild,accounting for 3.71％ of all the children examined,21 cases were mild,accounting for 0.88％ of all the children examined,7 cases were moderate,accounting for 0.29％ of all the children examined,and no severe cases were found.A total of 68 adults over 25 years old were examined for skeletal fluorosis.Twelve patients were found to have clinical skeletal fluorosis.The detection rate was 17.65％.X-ray radiography was used to examine 46 people,and 1 case was detected,the detection rate was 2.17％.Clinical and X ray examinations showed no more than moderate patients.Fifty samples of adult urine were tested,the level of urine fluorine was 0.72 mg/L.Conclusions Water improvement project is working well in Huzhu County,and no serious children with dental fluorosis and adult skeletal fluorosis are detected.The prevention and control measures of drinking-water-bornefluorosis has achieved remarkable results in Huzhu County.We will continue to strengthen water improvement project and disease monitoring,and thoroughly control the prevalence of drinking water fluorosis.

Objective:To observe and evaluate the effect of health education on drinking brick-tea type fluorosis in Zhiduo County, Qinghai Province, so as to provide basis for further formulating health education strategies.Methods:From April 2019 to April 2020, according to the historical prevalence of drinking brick-tea type fluorosis in Zhiduo County, Qinghai Province, 3 townships (towns) were selected to carry out the health education activities on drinking brick-tea type fluorosis for students of grade 4 - 6, village doctors, adults and monks in each township (town). We carried out a one-year publicity on the prevention and treatment of drinking brick-tea type fluorosis, distributed health education materials and organized health education activities. Before and after the intervention, we conducted a questionnaire survey on health education among the target population (grade 4 - 6 students, village doctors, adults and monks), to evaluate the awareness rate and behavior formation rate of fluorosis prevention and control, and to evaluate the intervention effect.Results:A total of 86 students of grade 4 - 6, 40 village doctors, 42 adults and 20 monks were investigated, after the intervention, the awareness rates of prevention and treatment of drinking brick-tea type fluorosis in grade 4 - 6 students, village doctors, adults and monks were 87.98% (227/258), 96.67% (116/120), 81.75% (103/126), 83.33% (50/60), respectively, which were significantly higher than those before the intervention [38.38% (76/198), 83.33% (100/120), 15.45% (19/123), 28.89% (13/45), P < 0.05]. After the intervention, the behavior formation rates of prevention and treatment of the drinking brick-tea type fluorosis in grade 4 - 6 students, village doctors, adults and monks were 74.42% (128/172), 72.50% (58/80), 52.38% (44/84), 60.00% (24/40), respectively, which were significantly higher than those before the intervention [14.39% (19/132), 38.75% (31/80), 3.66% (3/82), 0(0/28), P < 0.05]. Conclusion:The comprehensive intervention measures based on health education can significantly improve the knowledge of local residents, and improve their bad drinking habits of drinking tea, which is of great significance to the prevention and treatment of drinking brick-tea type fluorosis.

Objective:To study the epidemic status of drinking tea type endemic fluorosis in Qinghai Province.Methods:In 2019, in counties (cities, districts, referred to as counties) that had the habit of drinking brick tea in 8 cities (prefectures) of Qinghai Province, epidemiological investigation of drinking tea type endemic fluorosis was carried out in villages. Ten households were randomly selected from each village, to investigate the demographic data of each household and the drinking situation of brick tea, residents' drinking water and brick tea samples were collected to determine the fluorine content, and calculate the daily per capita tea fluorine intake. At the same time, skeletal fluorosis was examined in all adults over 25 years old, and dental fluorosis was examined in all children aged 8 to 12 years old in survey sites. The content of fluorine in tea and water was detected by ion selective electrode method; the diagnosis of skeletal fluorosis was based on "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS 192-2008), the diagnosis of dental fluorosis was based on "Diagnosis of Dental Fluorosis" (WS/T 208-2011).Results:The mean (range) of fluorine of the 3 602 water samples was 0.31 (0.20 - 1.00) mg/L. The geometric mean (range) of fluorine of the 31 067 brick tea samples was 646 (40 - 2 295) mg/kg, the fluorine content of the brick tea ≤300 mg/kg accounted for 7.80% (2 422/31 067) of the total samples. The proportion of drinking Fu brick-tea was 89.97% (27 952/31 067); and the daily per capita tea fluorine intake was 1.93 mg, the daily per capita tea fluorine intake in Guoluo, Yushu and Hainan prefectures were higher than the health standard (3.50 mg). The detection rate of skeletal fluorosis in adults was 0.16% (2 357/1 484 907), Yushu Prefecture was the highest [29.23% (592/2 025)], followed by Guoluo Prefecture, which was 8.21% (771/9 393). The detection rate of dental fluorosis in children was 4.79% (8 076/168 623), Yushu Prefecture was the highest [32.61% (1 562/4 790)].Conclusion:Drinking tea type endemic fluorosis is prevalent in Qinghai Province, with obvious regional characteristics, covering a large population. The disease is relatively popular in Yushu Prefecture and Guoluo Prefecture.

Objective:To find out the variety, producing area, fluorine content of brick tea in Qinghai Province, and the drinking situation of brick tea among people, so as to provide basis for preventing and curing endemic fluorosis of drinking tea type.Methods:From April to November 2019, according to historical data, in 3 066 administrative villages in 39 counties (cities, districts, hereinafter referred to as counties) in Qinghai Province that had the habit of drinking brick tea, 10 families were randomly selected from each village to investigate the demographic data of each family and the drinking situation of brick tea, to collect brick tea samples to determine the fluorine content, and to calculate the daily average brick tea fluorine intake of the population > 16 years old.Results:A total of 31 067 brick tea samples were collected. The main brick tea consumed by the residents in the whole province was Fu brick tea, accounting for 89.97% (27 952/31 067), followed by Kang brick tea [5.12% (1 592/31 067)], Green brick tea [2.29% (710/31 067)], Black brick tea [1.85% (574/31 067)], and golden tip, mosaic, black wool and other brick tea [0.77% (239/31 067)]. There were 523 brands of brick tea in circulation in the province, among which there were 410 brands produced in Hunan Province, 26 brands in Sichuan Province, 11 brands in Hubei Province, and 76 brands in Henan Province and other provinces. The average content of fluorine in brick tea was 646.1 mg/kg, which ranged from 40.0 to 2 295.0 mg/kg. Brick tea with fluorine content ≤300 mg/kg accounted for 7.80% (2 422/31 067) of all samples. The annual average consumption of brick tea by population > 16 years old was 1.09 (0.35 - 7.40) kg, and the daily average brick tea fluorine intake was 1.93 (0.39 - 18.64) mg. There were 15 counties and 486 administrative villages in which the daily average brick tea fluorine intake exceeded the national standard (3.5 mg).Conclusion:The main brick tea in circulation in Qinghai Province is Fu brick tea, which has high fluorine content and is harmful to people, and prevention and control measures should be taken as soon as possible.

Objective:To observe the changes of serum C-terminal peptide of type Ⅰ collagen (CTX-1) and N-terminal lengthening peptide of type Ⅰ collagen (P1NP) in adult patients with skeletal fluorosis in the tea-drinking-borne endemic fluorosis area in Qinghai Province, and to find sensitive indicators for diagnosis of skeletal fluorosis.Methods:From April to August 2019, a case-control study was carried out in tea-drinking-borne endemic fluorosis area in Zhiduo County, Yushu Tibetan Autonomous Prefecture, and Gangcha County, Haibei Tibetan Autonomous Prefecture of Qinghai Province. According to the Diagnostic Standard for Endemic Skeletal Fluorosis (WS/T 192-2008), the clinical diagnosis and X-ray examination of skeletal fluorosis were carried out for permanent residents ≥25 years old and living for more than 10 years in the area, combined with face-to-face inquiry and investigation of past disease history, lifestyle and clinical manifestations. The patients with skeletal fluorosis and healthy people were selected as skeletal fluorosis group and control group, respectively. Randomized urine samples and fasting venous blood from the two groups were collected. The content of fluoride in urine was determined by ion selective electrode method, and the contents of CTX-1 and P1NP in serum were determined by enzyme-linked immunosorbent assay (ELISA).Results:A total of 127 people in the disease area were investigated, including 63 cases in skeletal fluorosis group and 64 cases in control group. There was no statistically significant difference in age and sex ratio between the two groups ( t = 0.42, χ 2 = 0.07, P > 0.05). The X-ray examination results showed that the patients with skeletal fluorosis were mainly mild, accounting for 71.43% (45/63); X-ray changes were mainly ossification of interosseous membrane and tendon. The urinary fluoride in control group and skeletal fluorosis group was 1.62 (1.12, 1.95) and 3.22 (2.38, 4.89) mg/L, respectively, with statistically significant difference between the two groups ( Z = 7.07, P < 0.001). The difference of serum CTX-1 and P1NP contents between the two groups was statistically significant ( Z = 2.00, 4.89, P < 0.05). Conclusions:The levels of serum CTX-1 and P1NP in patients with skeletal fluorosis are higher than those in healthy people. Serum CTX-1 and P1NP may be used as sensitive indicators for diagnosis of skeletal fluorosis.