In order to solve some limitations and difficulties in guiding the practical work of determination and classification of the areas of high water iodine and the endemic areas of iodine excess goiter (GB/T 19380-2003),we carried out the revision on the original standard from 5 aspects such as the name,normative reference file,the definition and the technical indicators of classification of high iodine areas and the endemic areas of iodine excess goiter,such as sampling method in appendix and so on,and worked out the standard of definition and demarcation of water-borne iodine-excess areas and iodine-excess endemic areas.Enforcement of the new standard will bring important social benefits.

The Arabidopsis thaliana glutathione S-transferases zeta class (AtGSTZ) is a multi-functional enzyme, which plays important role in cellular metabolism and environmental purification. Error-prone PCR and cycles of DNA shuffling were used to construct a mutagenesis library of AtGSTZ. The screening of the resultant libraries was carried out by a pH indicator dye-based colorimetric assay. Nine mutants which enhanced the dichloroacetic acid dechlorination activity were obtained. Among them, NN23 contained 25 amino acid substitutions with the activity improving 120%, whereas NN20 contained 24 amino acid substitutions with the activity improving 102%. EC1 contained 2 amino acid substitutions with the activity improving 47%. The rest 6 mutants contained one amino acid substitution with their activity increasing from 9% to 60%. The enzymatic characterization showed that all the evolved enzymes increased their catalytic efficiencies towards dichloroacetic acid and binding affinity towards glutathione whereas some of them increased the renaturability. However there is no obvious change in their thermostability. Based on these data, functional residues related to catalysis and refolding of AtGSTZ were discussed.

AIM:To construct nine novel L-asparaginase mutants and study their enzyme-activity.METHODS:The mutants were constructed using overlap extension PCR according to the principle of alanine-scanning mutagenesis. The enzyme-activity was detected by Nessler's method. RESULTS:The DNA sequencing showed that the mutagenesis was consistent with the theoretical prediction. The enzyme-activity assay demonstrated that each mutant possessed enzyme activity equal to the original enzyme. CONCLUSION:Through gene modification,epitop of L-asparaginase was changed without activity loss.These results provide foundation for further study of the structure-function relationship of L-asparaginase.

Objective Toinvestigate iodine nutrition,thyroid volume and function of children and women in high water iodine areas,and to discuss the cut-off point of water iodine level where should be defined as iodine excessive areas.Methods In iodine excessive townships in Xiaodian and Qingxu Counties of Taiyuan City,all villages were divided to 4 groups according to the concentration of iodine in drinking water:50 ～ ＜ 100 μg/L (Group A),100 ～ ＜ 150 μg/L(Group B),150 ～ ＜ 300 μg/L(Group C) and ≥300 μg/L(Group D),and 2-3 villages were randomly selected from each group as investigation sites.Two hundred children aged 8-10 and 60 women (20 pregnant women,20 breasffeeding women and 20 women of childbearing age,respectively) were sampled.Drinking water,edible salt and once random urine samples for each studying object were collected,and the iodine content was measured.The goiter volume of children aged 8-10 and triiodothyronine (FT3),free thyroxine (FT4),thyroid stimulating hormone (TSH) of women were determined.Results Totally 708 children and 236 women were selected as respondents who ate non-iodized salt.Iodine content in drinking water of children in groups A,B,C and D was 73.8,144.7,258.5 and 501.0 μg/L,respectively,and that of women was 73.8,144.7,242.7 and 485.9 μg/L,respectively.Median urinary iodine of children in groups A,B,C and D was 274.3,312.8,455.6 and 793.5 μg/L,respectively,and that of women was 238.3,235.2,371.6 and 641.6 μg/L,respectively.The median urinary iodine of children and women increased with increasing water content of iodine.The goiter rate of children was 5.6％ (11/196),13.2％ (25/189),12.6％ (20/158) and 10.9％ (18/165) for each group,respectively,which also increased with increasing water content of iodine.In groups A,C and D,the FT3 and FT4 levels[pmol/L:(3.70 ± 0.59),(14.01 ± 2.44); (3.43 ± 0.57),(12.87 ± 2.12); (3.42 ± 0.47),(12.66 ±1.78)] in pregnant woman were lower than those in breasffeeding women[pmol/L:(4.26 ± ±0.57),(14.73 ± 2.36;(4.28 ± 0.40),(14.77 ± 1.19); (4.36 ± 0.65),(15.66 ± 2.84)] and women of childbearing age[pmol/L:(4.80 ±0.50),(17.47 ± 2.11); (4.21 ± 0.48),(15.83 ± 1.64); (4.26 ± 0.52),(15.53 ± 1.81)].With increasing water content of iodine,FT4 level was decreasing and TSH level was increasing gradually in women.Conclusions When water iodine exceeds 100 μg/L,goiter rate of children has increased significantly.Iodine excessive women and children have appeared one after another with increasing water content of iodine.Women incidence of subclinical hypothyroidism risk has increased with increasing water content of iodine,and the detection rate of subclinical hypothyroidism is significantly higher when water iodine is higher than 300 μg/L.The cut-off point of iodine excessive areas should be descend from 150 μg/L to 100 μg/L.

Objective:To investigate the iodine nutrition level of pregnant women in Shanxi Province, and to provide basis for guiding pregnant women to supplement iodine scientifically.Methods:From 2016 to 2018, 40, 80 and 118 counties were selected in Shanxi Province. In each county, 5 townships were selected according to the east, west, south, north and middle orientation, 20 pregnant women were selected from each township. Urine and salt samples of pregnant women were collected, and urinary iodine and salt iodine contents were determined, respectively.Results:From 2016 to 2018, 3 590, 7 907, and 11 750 salt samples were collected from pregnant women's homes. The medians salt iodine were 23.80, 23.70, 23.25 mg/kg, and the qualified iodized salt consumption rates were 91.06% (3 269/3 590), 90.06% (7 121/7 907), 92.21% (10 835/11 750), and the coverage rate of iodized salts were 97.72% (3 508/3 590), 97.00% (7 670/7 907), 98.53% (11 577/11 750), and the qualified rate of iodized salts was 93.19% (3 269/3 508), 92.84% (7 121/7 670), 93.59% (10 835/11 577), respectively. There were statistically significant differences in the median salt iodine and qualified iodized salt consumption rate among pregnant women in different years ( H = 99.915, χ 2 = 27.988, P < 0.05). Totally 3 902, 7 892 and 11 745 urine samples of pregnant women were tested, and the medians (quartiles) urinary iodine were 174.20 (114.00, 251.08), 180.70 (117.13, 258.58) and 179.40 (115.90, 249.00) μg/L, respectively. There was statistically significant difference in the median urinary iodine among pregnant women in different years ( H = 12.368, P < 0.05). From 2016 to 2018, counties with appropriate levels of iodine accounted for 70.00% (28/40), 73.75% (59/80) and 70.34% (83/118), respectively. The medians (quartiles) urinary iodine levels of pregnant women in the early, middle and late stages of pregnancy ( n = 2 225, 5 727, 3 793) in 2018 were 177.62 (117.28, 257.23), 178.21 (117.40, 248.40) and 172.70 (112.98, 245.70) μg/L, respectively, and there was statistical by significant difference in urinary iodine levels among pregnant women in different pregnancy periods ( H = 11.077, P < 0.05). Conclusions:From 2016 to 2018, the iodine nutrition level of pregnant women in Shanxi Province is generally in an appropriate state, iodine deficiency or more than appropriate still exists in some areas. We should continue to carry out monitoring work, and focus on strengthening health education for pregnant women, and guide them to supplement iodine scientifically and accurately.

Objective To obtain the protein interacting with inhibitor of differentiation1′(Id1′). Methods The recombinant bait plasmid pHybLex/Zeo Id1′ was constructed and transformed into yeast strain EGY48/ pSH18 34 to test pHybLex/Zeo Id1′ for non specific activation. Adult human lung cDNA libraries were screened to obtain true positive library plasmid. The true positive library clone was obtained by sequencing and basic local alignment sequence tool (BLAST). Results The recombinant bait vector, named as pHybLex/Zeo Id1′, was confirmed by sequencing. pHybLex/Zeo Id1′ was transformed into yeast strain EGY48/pSH18 34 and the transformants had no autonomously activated reporter genes. One true positive clone, obtained by screening of the adult human lung cDNA libraries, was confirmed to be Fyn by sequencing and BLAST. Conclusion Id1′ can interact with Fyn.

Objective To investigate the influence of cerebral lymphatic blockade (CLB) on apoptosis of hippocampal neurons after subarachnoid hemorrhage (SAH) in rats. Methods Healthy adult Wistar rats were randomly assigned to normal control group,SAH group and SAH + CLB group. SAH model was induced by double injection of autologous blood into the cistema magna. On day 3 after second injection, hippocampal cell shape structure of each group were determined by hematoxylin-eosin staining (HE) and propidium iodide (PI) staining. Terminal-deoxynucleotidy transferase mediated nick end labeling (TUNEL) fluorescent was used to determine the situ apoptosis. Immunohistochemistry was conducted to study the expression of caspase-3 and Bcl-2 in hippocampal neurons. Results (1) HE staining and PI staining showed the hippocampal neurons of SAH rats were partly shrink,and nuclei showed wavy or folded seam-like,some crescent-shaped; the hippocampal neurons in SAH + CLB group distributed sparsely,nuclear fragmentation,apoptotic bodies could be seen,surrounded by vacuole formation, Compared with the SAH group, the number of apoptotic cells in SAH + CLB group was significantly increased(the number of apoptotic cells: 0.71 ±0.05,25.36 ±4. 02,37. 82 ±5.93, P<0.01). (2) The fluorescence intensity of positive cells by TUNEL stain in SAH group and SAH + CLB group was higher than in normal control group,while the SAH + CLB group was significantly higher than the SAH group (the fluorescence intensity: 0.19 ±0.03,1.70 ±0.37,2.54±0.53, P<0.01). (3) The fluorescence intensity of caspase-3 in SAH group and SAH + CLB group was higher than the normal control group, while the SAH + CLB group was significantly higher than the SAH group (the fluorescence intensity: 0.14 ±0.03,2.45 ±0.49,2.96 ±0.44, P<0.01). (4) The fluorescence intensity of Bcl-2 in SAH group and SAH + CLB group was higher than the normal control group, while the SAH + CLB group was significantly lower than the SAH group(the fluorescence intensity: 0.58 ±0.08, 3.40 ±0.61,2.67 ±0.44, P<0.01). Conclusion Cerebral lymphatic blockade induce the apoptosis of hipp-ocampal neurons in rats after SAH,which mechanism may be related to high expression of caspase-3 and low ex-pression of Bcl-2.

Objective To investigate the nutritional status of iodine among residents and the goiter disease of children in high water iodine areas in Shanxi Province,and to provide a scientific basis for establishment of longlasting control strategies and measures.Methods In 2012,according to the historical monitoring data,in the 10 high water iodine counties (citys,districts),one town was selected based on its location (east,west,south,north and middle) in each county (citie,district).In county (city,district) with 5 townships or less,all townships were selected.Four villages were selected in each township and fifteen residents in each village were selected to test salt iodine level.In five high water iodine counties (city,district),one or two high water iodine villages were selected,water samples were collected and the iodine content was measured; one hundred students aged 8-10 years old were chosen to examine their thyroid size.Thirty children were chosen from above students to collect their urine samples and to determine the iodine content.Results In 10 high water iodine counties (citys,districts),1 680 salt samples were tested.The rate of non-iodized salt was 85.2％ (1 432/1 680); in six villages of five high water iodine counties (citys,districts),the median of urinary iodine of 256 children aged 8 to 10 was 487.2 μg/L; in three villages which had improved the quality of water,the median of urinary iodine was 271.0 μg/L; other three villages which had not improved the quality of water,the median of urinary iodine was 692.6 μg/L.In those villages which had not improved the quality of water,urinary iodine of children ≥300 μg/L accounted for 85.8％ (139/162); in those villages which had improved the quality of water,high urinary iodine of children accounted for 41.5％ (39/94),and the difference was statistically significant (x2 =53.06,P ＜ 0.05).The thyroid was investigated among 591 children aged 8-10 years old,and the goiter rate was 6.6％(39/591).In those villages which had improved the quality of water,the goiter rate of children was 3.8％ (11/291),but in villages which had not improved the quality of water,the goiter rate of children was 9.3％ (28/300),and the difference was statistically significant (x2 =6.52,P ＜ 0.05).Conclusion The iodine nutrition level of residents in high water iodine areas in Shanxi is excessive,children's goiter disease has not been effectively controlled; water improvement to reduce iodine is the basic way to control the disease of high iodine.

Objective To analyze the effect of adjustments of control strategy on epidemic trend of iodine deficiency disorders (IDD) in Shanxi Province after universal salt iodization (USI),and to provide basis for timely adopting targeted control countermeasure and scientifically adjusting intervention strategy.Methods A method of retrospective analysis was performed to collect data from IDD surveillance at national or province levels after 1995,and from iodized salt surveillance of the province after 2004.According to the statistics and analysis of children's goiter rate,median urinary,median and mean of salt iodine,coverage rate of iodized salt,qualified rate of iodized salt,consumption rate of qualified iodized salt and their relationship.Results Since 1995,the children's goiter rate by palpation and B-ultrasound showed a steady descending trend.The median salt iodine,mean salt iodine and children's median urinary iodine showed a trend of rise→decline→stable→decline.Namely:The three indicators began to rise year by year from 1995 (29.1 mg/kg,31.7 ± 15.0 mg/kg,199.3 μg/L),in 1999 (48.7 mg/kg,53.4 ± 29.4 mg/kg,407.5 μg/L) reached its climax; and then decreased,in 2001 (34.7 mg/kg,36.2 ± 11.9 mg/kg,282.1 μg/L)stoped; which were basically stable from 2001 to 2011; since 2013 (26.0 mg/kg,26.5 ± 6.3 mg/kg,192.0 μg/L),a significant decline began.The rate and edible rate of qualified iodized salt showed a trend of decline→rise→stable.Two indexes began to decline circuitously from 1995 (72.61％,68.25％),and dipped to a low point in 1999 (44.80％,43.67％); then began to rise,until 2002 (94.73％,91.80％) reached basic stability; and remained steady from 2002 to 2013.Conclusions Following the process of prevention and treatment of IDD for more than 30 years in Shanxi Province,with the depth understanding of the range of adequate iodine nutrition,according to the monitoringfeedback mechanism,the strategy of salt iodization has been adjusted several times,the target of continuous elimination of IDD has achieved since 2000 and the levels of iodine nutrition in population are more reasonable.Salt iodization strategy should continue to adhere to.

Objective To explore the effects of Kangshuaiyizhi capsule on ChAT/AchE and NE, DA and 5-HT levels in the brain tissue of aging model rats, and explore its effect of protecting cerebral function. Methods Totally 72 rats were randomly divided into normal group and model group. The subacutely aging model rats were made by injecting D-gal (0.125 g/kg) into abdominal cavity continually, then aging rats were divided by random number table into model group, Naofukang group and Kangshuaiyizhi high-, low-dose group. After intervented with correspongding drugs for 60 days, activity of ChAT and AchE, cerebral cortex NE, DA, and 5-HT levels were detected. Results Activity of AchE was much higher (P<0.05), but level of ChAT, NE, DA and 5-HT in model group were significantly downregulated compared with normal group (P<0.05). After treated with Kangshuaiyizhi capsule, activity of AchE was downregulated, ChAT, NE, DA and 5-HT levels were significantly upregulated (P <0.01, P <0.05). Conclusion Kangshuaiyizhi capsule can regulate cholinergic and monoamine neurotransmitter levels in the brain tissue of aging model rats, and play a very important role in protecting cerebral function.