Objective To investigate the expression of microRNA-29b (miR-29b) in gastric cancer cell line GC9811 and high peritoneal metastatic gastric cancer cell line GC9811-P and its effect on invasion,proliferation and apoptosis.Methods The relative quantitative expression of miR-29b was detected by quantitative realtime polymerase chain reaction(qRT PCR).GC9811 cells were divided into three groups,miRNA down-regulated and transfected with lentiviruses LV-miR-29b inhibitor group,negative control group with negative transfection,and untransfected blank control group.GC9811-P cells were divided into three groups,miR-29b up regulated and transfected with lentiviruses LV miR 29b group,negative control with negative transfection group,and untransfected blank control group.The cell invasion ability was detected with Transwell assay,the cell proliferation ability was measured by methyl-thiazolyl tetrazolium (MTT) test,the colony forming ability was determined by plate colony formation assay,and the apoptosis was tested by flow cytometry.The expressions of miR 29b and secreted protein,acidic and rich in cystenie (SPARC) in gastric cell line GC9811-P,GC9811,MKN28M,MKN28NM and normal gastric cell line GES were determined by qRT-PCR,and the correlation was analyzed.Two independent samples t test or SNK-q test was performed for mean comparison between two groups,and one way analysis of variance was used for mean comparison among three groups.Results The relative quantitative expression of miR-29b inGC9811-P (0.21±-0.04) was significantly lower than that of GC9811 (1.00±0.03,t 28.140,P＜ 0.01).After GC9811 cells transfected with lentiviruses LV-miR-29b inhibitor,the expression of miR-29b (0.21±0.04) was significantly lower than that of control group (0.89±0.07) and blank control group (1.00±0.04,q 12.76,14.73,both P＜0.01).Compared with negative group,the transmembrane cell number and the clonality of miRNA up-regulated group raised(274.33± 9.03 vs 110.67 ± 13.69,t=9.981,P＜0.01;131.33±4.91 vs69.67±2.33,t 11.340,P＜0.01),and the results of MTT test also shows the proliferation was increased.After GC9811-P cells transfected with LV-miR-29b,the expression of miR 29b (4.08±0.20) was significantly higher than that of negative control group (1.15±0.05) and blank control group (1.00±0.10,q=21.73,22.81,both P＜0.01).Compared with negative group,the transmembrane cell number and the clonality of miRNA up regulated group reduced (51.33±5.55 vs 104.00±6.24,t 6.305,P＜0.01; 48.00±5.51 vs 113.33±5.17,t 11.340,P＜0.01),and the results of MTT test also shows the proliferation was weakened.Apoptosis assays demonstrated miR-29b promoted apoptosis; however,the difference was not statistically significant.The expression of miR-29b was negatively correlated with SPARC mRNA in gastric cancer cells (r=-0.97,P=0.03).Conclusions The low expression of miR-29b in high peritoneal metastatic gastric cancer cell inhibited the ability of invasion and proliferation.MiR-29b might be a new target of inhibiting peritoneal metastasis in gastric cancer.

Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction (qRT-PCR ). GC9811 cell line was transfected byendogenous synthetic analog mimic and its homologous negative control of miRNA-181a-5p,which were considered as up-regulated group and its control group respectively;GC9811-P were transfected by miRNA inhibitor and its homologous negative control of miRNA-181a-5p,which were considered as down-regulated group and its control group respectively.After miRNA-181a-5p was up or down-regulated,cell proliferation,migration and apoptosis capabilities of gastric cells were detected by matrix thiazolyl tetrazollium (MTT)assay,cloning-forming assay,Transwell migration test,wound healing assays and apoptosis test.After miRNA-181a-5p was up or down regulated,the changes of matrix metalloproteinase (MMP)14 expression were determined by Western blot.Independent sample t test was performed for mean comparison between samples and chi square test was used for rate comparison.Results The results of qRT-PCR showed the relative expression quantity of miRNA-181a-5p in GC9811 was 1 .000 00 ± 0.021 26 and in GC9811-P was 3.175 61 ±0.106 76,and the difference was statistically significant (t =34.620,P <0.01 ).The results of MTT assay indicated that the cell proliferation rate of up-regulated group was higher than that of up-regulated control group,and that of down-regulated group was lower than down-regulated control group.The cloning-forming assay demonstrated that the number of clone forming and clone forming rate of up-regulated group,up-regulated control group,down-regulated group and down-regulated control group were 234.00±10.12 and 46.8%,93.00±9.61 and 18.6%,51 .00 ±7.96 and 10.2%,99.00±8.05 and 19.8%,respectively.The differences between up-regulated group,down-regulated group and their control group were statistically significant (t = 17.500,7.344,χ2 = 12.27, 9.51 ,all P <0.01).The results of Transwell migration experiment showed the number of cells migrated through membrane hole of up-regulated group was 164.00±19.31 ,and which was higher than that of up-regulated control group (87.00±23.04,t=4.436,P <0.05);that of down-regulated group was 157.00± 11 .50,and which was lower than that of down-regulated control group (234.00 ±12.12,t =7.982,P <0.05).The result of wound healing assays indicated that the rate of migration distance of up-regulated group,up-regulated control group,down-regulated group and down-regulated control group were 2.09 ± 0.18,1 .27 ±0.23,1 .15 ±0.15 and 1 .67 ±0.12,respectively.The differences between up-regulated group,down-regulated group and their control group were statistically significant (t =4.863 and 4.689, both P <0.05).The results of apoptosis experiments demonstrated that the apoptosis rate of up-regulated group,up-regulated control group,down-regulated group and down-regulated control group were (6.10± 1 .02 )%,(9.10 ± 2.13 )%,(12.70 ± 1 .23 )%,(8.70 ± 2.54 )%,respectively,and there was no statistically significant difference between up-regulated group,down-regulated group and their control group (both P *0.05).The results of Western blot showed that the grey value of up-regulated group was 561 .881 ±35 .740,which was higher than that of up-regulated control group (275 .784±23.520);that of down-regulated group was 579.565 ±37.950,which was lower than that of down-regulated control group (1 312.760±51 .270),and the differnces were statistically significant (t =11 .580 and 19.910,both P <0.01).Conclusion miRNA-181a-5p highly expresses in peritoneal high metastasis gastric cancer cell line GC9811-P and promoted the proliferation and migration of gastric cancer cell line GC9811 and GC9811-P with a tendency to suppress apoptosis.

Objective To investigate the effects of selenium and iodine deficiency on the contractibility of rat thoracic aorta and probe into its mechanisms.Methods Isolated vascular methods were used to study the effects in different groups.Results The contraction of rat thoracic aorta of the groups with selenium and iodine deficiency was less than that of the normals(P(0.05));sodium selenite(0.1mol/L) had no effect on rat aortic contraction,but was significantly synergic with the contraction induced by KCl combined with NE(P

Objective To investigate the epidemic status of endemic fluorosis in Shandong Province,Jining City,and to provide a basis for prevention and control of the disease.Methods Based on Shandong Provincial Project Technical Solutions for Endemic Fluorosis,Rencheng,Jinxiang,Yutai,Jiaxiang and Liangshan Counties in Jining were selected as monitoring sites.According to the illness situation of mild,moderate or serious districts,one village was selected as a major survey site from each county(district).There were a total of 15 such villages selected.Survey content included drinking water fluorine level; dental fluorosis of children,adults' clinical skeletal fluorosis and urinary fluorine levels; water and urinary fluoride content were determined by the method of fluoride ion selective electrode; dental fluorosis of children was diagnosed by Deans method and clinical diagnosis was based on the Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008).Results Sixty-one water samples from 15 villages of five counties (districts) were tested.Fluoride levels of 9 out of the 61 samples were exceeded the national standard (＞ 1.0 mg/L),and the rate was 14.75％; 1 sample ＞ 2.0 mg/L,and the maximum water fluoride was 2.25 mg/L.Seven hundred and seventeen people's real time urinary fluoride was detected in the 15 villages,including 420 children and 297 adults,and the geometric mean were 1.53 and 1.69 mg/L,respectively.Clinical examination of 755 children aged 8 to 12 showed that the detection rate of dental fluorosis was 26.89％ (203/755); defect rate was 9.12％(29/755) and dental fluorosis index weres 0.65.The detection rate of clinical skeletal fluorosis of 11 565 adults was 4.76％(550/11 565),including 303 moderate or serious cases.Conclusions The situation of excessive water fluorine in outside environment in Jining City has been controlled at a certain degree; groups urinary fluoride level is closed to the normal upper limit; the prevalence of dental fluorosis or skeletal fluorosis has been suppressed at a certain degree,therefore,the results of control should be further consolidated and expanded,in order to completely eliminate the fluoride hazard.

Objective To analyze the assessment results of external quality control and network operation of urinary iodine in local laboratories of Shandong Province,to evaluate the ability of consistent analysis; and to provide a reliable laboratory quality assurance for epidemiological surveillance and control of iodine deficiency disorders (IDD) and reliable decision-making.Methods Z-scores of the assessment results of external quality control of urinary iodine in local laboratories of Shandong Province were analyzed from 2000-2013.Results The feedback rate of urinary iodine in local laboratories of ShandongProvince was 100.0％ from 2000-2013; the qualified rate was 100.0％(14/14),100.0％(14/14),92.9％(13/14),100.0％(14/14),92.9％(13/14),100.0％ (14/14),100.0％ (14/14),100.0％ (14/14),92.9％ (13/14),100.0％ (17/17),100.0％ (17/17),94.1％ (16/17),100.0％ (17/17),and 100.0％ (17/17),respectively.The total qualified rate of Z-scores between Zs in local laboratories was 100.0％ (214/214)and within Zs was 98.6％ (211/214).Conclusion The test abilities of urinary iodine in local laboratories of Shandong Province are steady; the overall results are satisfactory,but some laboratories need to improve.

In order to study genetic variation diversity of porcine circovirus type 2 (PCV2) strains in Shanxi,the genomic sequences of nine PCV2 strains including SXQX,SXCZ,SXTY2,SXJC,SXJX,SXLL,SXPY,SXPG and SXXY recently isolated from some areas of Shanxi from 2013 to 2016,was cloned,sequenced and received by GenBank.The amplified PCV2 genomic sequences,ORF2 sequences and Cap protein amino acid of these nine strains were analysed and compared with those of published 28 PCV2 strains by DNAStar,drawing phylogenetic tree.The results showed that the genomic sequences of SXJX,SXJC and SXXY PCV2 strains were 1 768 bp,and the others were 1 767 bp,which accounted for 33％ and 67％,respectively.The homologies of nucleotide sequences of the nine strains were 94.7％-99.8％,the homologies of nucleotide sequences of the nine strains with the 28 isolates from different regions of the world PCV strain were 93.9％-99.9％,and the homologies of nucleotide sequences of the nine strains with the domestic vaccine strains were 95.1％-99.8％.The phylogenetic analysed that SXJX,SXJC and SXXY belonged to genotype PCV-2D,SXLL,SXPY and SXCZ belonged to genotype PCV-1C,and SXTY14,SXPG and SXQX belonged to genotype PCV-1A/1B.Thus it proved that the epidemic strain of PCV2 was mainly PCV-2b in Shanxi.The homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains were 90.0％-100.0％ and 87.1 ％-100.0％ respectively,the homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains with the 28 isolates from different regions of the world PCV strain were 87.6％-100.0％ and 84.1％-100.0％ respectively,and the homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains with the domestic vaccine strains were 91.0％-100.0％ and 89.3％-100.0％ respectively.The Cap amino acids of SXQX,SXJX,SXTY14,SXPG,SXJC and SXXY PCV2 were 233,ORF2 of SXQX,SXTY14 and SXPG located at 1 033-1 734 bp,ORF2 of SXXY,SXJX and SXJC located at 1 033-1 734 bp,and the Cap amino acids of SXCZ,SXLL and SXPY PCV2 were 234,ORF2 of them located at 1 030-1 734 bp,in addition,the positions of 1 030-1 734 bp were more three bases TCA than other ORF2 genome sequence of 1 767 bp,resulting in increasing a K (Lys) of amino acid sequencein at the 234 position.Also Cap protein of 9 PCV2 strains showed more amino acid variation in addition to the only high-ly conserved glycosylation sites (NYS) (pp.143-145 amino acid).It provided theoretical basis for the PCV2 immune prevention of research in Shanxi,and the data of basic theory of molecular pathogenesis of PCV2.

Objective:To determine the prevalence and risk factors for age-related macular degeneration (AMD) in Tibetan of plateau area, and to establish a nomogram prediction model.Methods:A cross-sectional study was carried out.The 40-year-old and older Tibetan subjects were enrolled by cluster random sampling from Xinghai County and Tongde County with an average altitude of 3 000 meters in Qinghai Province.Questionnaire survey, routine ophthalmic examinations, direct fundus photography and health education were carried out among the subjects.The questionnaire items included age, gender, smoking, drinking, living duration in plateau region, daily sunshine duration, diabetes, hypertension and hyperlipidemia.A χ2 test was used to screen significantly factors, and risk factors for AMD were analyzed by logistic regression analysis.The nomogram plot was drawn by R software to establish a predicting model.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Qinghai Provincial People's Hospital (No.2017-21). Written informed consent was obtained from each subject prior to any medical examination. Results:A total of 1 223 subjects were enrolled, and AMD were detected in 190 subjects with an prevalence of 15.54%.The risk factors for AMD were old age, living duration in plateau region, sunshine duration, smoking, hypertension and hyperlipidemia.Age was the first one of risk factors for AMD ( OR: 20.183, 95% CI: 9.536-29.805, P<0.001), followed by sunshine duration ( OR: 3.785, 95% CI: 1.581-8.721, P<0.001). The nomogram prediction model showed that when the questionnaire score was >150, the probability of suffering AMD was 50%.The decision curve showed that when the threshold probability was >25%, the predicted probability of AMD was the same as the actual probability. Conclusions:Pathogesis of AMD in Tibet is associated with old age, living duration in plateau area, sunshine duration, smoking, hypertension and hyperlipidemia.

OBJECTIVE To determine speech perception,tone perception and the subjective preferences of mandarin-speaking adults who received the fine structure processing(FSP) coding strategy,at upgrade from the continuous interleaved sampling(CIS+) coding strategy.METHODS Thirteen mandarin-speaking subjects were tested at switch-over from CIS+ to FSP coding strategy approximately 1-,2-,and 3-month after switch-over with the mandarin hearing in noise test(M-HINT),the mandarin tone identification in noise test(M-TINT),and a visual analogue scale assessing sound and speech assessment(SSA).RESULTS Tone perception improved significantly over time with the FSP coding strategy.There were no significant differences in the MHINT over time,nor when compared to the CIS+ coding strategy.Subjects rated the FSP coding strategy with the OPUS 2 as significantly more "full" and "rich" than with the CIS+ coding strategy after 3-months.CONCLUSION Mandarin-speaking adults can use the FSP coding strategy as well as CIS+ coding strategy.Loss in performance was not observed after switch-over.Tone identification improves over time with the FSP coding strategy,which is beneficial to tonal-language users.

Objective To evaluate the testing ability of iodine in the disease control and prevention institutes at all levels in Shandong,and to raise their testing ability.Methods The testing ability of salt iodine,water iodine and urinary iodine of iodine deficiency disorders laboratories at provincial,prefectural and county levels in Shandong in 2016 was evaluated.The testing results of salt iodine were evaluated by using reference value ± uncertainty.The testing results of water iodine and urinary iodine of all the participatory laboratories were evaluated by using standard Z score generated from laboratories participated in the examination.Results One provincial and 17 prefectural salt iodine,water iodine and urinary iodine laboratories and 30 county level salt iodine laboratories took part in the national examination.Both the feedback rate and qualified rate of the testing results were 100％.The 65 urinary iodine laboratories at the county level took part in the provincial examination.The feedback rate was 92.3％ (60/65),and the qualified rate was 75.4％ (49/65).Conclusions The results of the national examination have showed that the testing ability at all levels of the laboratory is maintained at a higher level;the results of the provincial examination have showed that the testing ability of most of the county level urinary iodine laboratories is relatively stable at a higher level.The testing ability of some county level urinary iodine laboratories is low.We should focus on strengthening the county level urinary iodine laboratory construction.

Objective To evaluate feasibility of a method for determining low and medium concentrations water iodine by an automatic iodine analyzer (hereinafter referred to as this method).Methods The low (0-10 μg/L)and medium (0-100 μg/L) concentrations water iodine were determined by an automatic iodine analyzer.Methodological experiments were carried out on this method,including standard curve linearity,detection limit,precision experiment,standard recovery experiment,and standard substance determination.Results The absolute values of the linear correlation coefficients of the low and medium concentrations standard curves were ＞ 0.999 0,and the detection limits were 0.32 and 2.60 μg/L,respectively.Precision:the coefficient of variations were all below 2％ (n =18) of low,medium and high iodine concentrations water samples in the range of low and medium concentrations water iodine,and the average recovery rates were 100.7％ and 101.1％ (n =6),respectively.The determination results of the water iodine standard substance GBW 09113d and GBW 09114d were (8.3 ± 0.3)and (61 ± 2) μg/L (n =6),respectively,which were within the given standard values range.Conclusions This method has higher precision and accuracy,less reagent consumption,less time consumption,and simple operation.It is convenient for large-scale detection,and has strong applicability.