Objective To establish a new and convenient assay for whole blood lead concentration. Methods Whole blood sample was digested by nitric acid-perchloric. Under the condition of pH 0.8～1.0, lead concentration was examined by Hydridegeneration Atomic Fluorescence Spectrometry.Results The method had good linearity when lead concentration were between 0 to 600 ?g/L, r =0.999. The within-run CV of the method for high, middle and low lead level were 4.1%, 3.6% and 3.5% respectively. The between-run CV were 4.9%, 4.6% and 4.7% respectively. The recovery rates were 105.4%, 98.5% and 94.8% respectively. The single-blind test result of series standard lead samples provided by Chinese Academy of Preventive Medicine were within the acceptable range. The detecting Limit of this method was 0.3 ng. There was no significant interference from main two-valence positiveion. The sample can be storage at lest for 1 months at 4℃. Samples of 869 children from Tianjin were tested by this method. The mean value was 91.9 ?g/L, median was 83.8 ?g/L, all data were within 23.1～386.3 ?g/L. The result showed out positive skewness distribution.Conclusion This method were sensitive, accurate, precise, quick and low-cost. It was suitable for use in clinical laboratories.

Adolescent ; Female ; Heart Septal Defects, Atrial ; complications ; therapy ; Humans ; Male ; Pulmonary Veins ; abnormalities ; Young Adult

Objective To observe endoplagmic reticulum stress in bone tissue of fluomsis rats and further explore the pathogenesis of skeletal fluorosis.Methods 48 Wistar rats were divided into 4 groups according to their body mass.The control and low.calcium group were fed with normal diet(0.79%calcium)and low-calcium diet(0.79%calcium)respectively,and both drank tap water(sodium fluoride concentrations＜1 mg/L).High fluoride and low.calcium plus high-fluoride groups were fed with normal diet(0.79%calcium)and low-calcium diet (0.79%calcium)respectively,and both drank tap water containing sodium fluoride(sodium fluoride concentrations 221 mg/L).During experimental period,rats were measured body mass once a week with a stand diet and water available ad libiturn.The experimental period was 3 months.The biochemical techniques were used to test the indicators of oxidative stress and ALP in seFum of fluorosis rats.The total RNA was extracted from the one side of the femur,and the transcription level of Bip,Xbp1,CHOP and PDI were investigated by reverse transcription polymerase chain reaction(RT-PCR).Results The level of MDA in serum of low-calcium plus high-fluoride group wag higher than that of the control[(14.74±3.11)μmol/L vs(10.15±1.96)μmol/L,P＜0.05];the activity of GPx was ma~edly higher in hish-fluoride group compared with the control[(3.87±0.41)×103 U/L vs(2.85± 0.55)×103 U/L,P＜0.05];the level of uric acid in sel'um was significantly lower both in high-fluoride group and low-calcium plus high-fluoride group compared with the respective control and the low-calcium group[(73.95± 9.52)μmol/L vs(110.43±25.48)μmol/L,(54.32±22.09)μmol/L vs(101.71±17.01)μmol/L,P＜0.05]. The activity of ALP wag obviously higher in low-calcium plus high-fluoride group compared with the control [(24.77±4.57)×103U/L vs (12.91±3.97)×103U/L,P＜0.01)].The mRNA expression of Bip/GAPDH in bone tissue was markedly higher in bone of high-fluoride group and low-calcium plus high-fluoride group compared with the control(1.38±0.24,1.35±0.12 vs 1.14±0.06,P ＜ 0.05). The expression of Xbp1/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control and the low-calcium group (1.48±0.20 vs 1.02±0.25,1.07±0.25,P ＜ 0.05 or ＜ 0.01);and CHOP/GAPDH in bone tissue significantly increased in low-calcium plus high-fluoride groups compared with the control(0.84±0.18 vs 0.52±0.07,P ＜ 0.05 ). Conclusions Accelerated osteogenetic action is seen in fluorosis rats,accompanied by oxidative stress and bone endoplasmic reticulum stress,which is likely involved in the pathogenesis of skeletal fluorosis.

Objective To study cervical central group (VI region) lymph node metastasis with papillary thyroid microcarcinoma and correlative influencing factors.Methods Clinical data of 215 papillary thyroid microcarcinoma patients undergoing surgery between Jan 2007 and Jan 2011 were analyzed retrospectively.Results All patients accepted bilateral thyroidectomy and bilateral cervical central group lymph node dissection.The total incidence of cervical central group lymph node metastasis was 36.7％ (79/215).Factors relating to cervical central group lymph node metastasis rate were:age (with one year elder,the likelyhood of lymph node metastasis was 0.935 times lesser),gender (the ratio of female to male was 0.202),the number of foci and the sum of the diameters of all lesions.Conclusions Patients with papillary thyroid microcarcinoma may suffer from lymph node metastasis of the cervical central group,and lymph node metastasis rate increases significantly in young or male patients,and when the lesions were multifocal or the sum of the tumor diameter ＞ 0.5 cm.

The effect of pH on the extracellularpolysaccharide synthesis by Aureobasidium pullulans was studied by addding CaCO 3and HCl.Cultivated in P2 liquid medium for 24h,pH dropped to 3.6 because of strongly producing acid.Under this low pH environment,further fermentation for 120h,only 5.9g/L of polysacharide was obtained.When grown in MP2 medium contaning0.5% CaCO 3,the pH was kept above 5.0 during 144 hours,production of polysaccharide increased to 31g/L. The detailed information of effects of controlled pH on polysaccharide production showed an optimal pH value 5.0 must be maintained through the fermentative period.

Objective To observe the expression of vascular endothelial growth factor(VEGF) mRNA and protein in fluoride(F~-) treated fibroblast(FB) of mice in planar(2D) and FBs populated collagen lattice(3D) culture systems and to further explore the effects of VEGF on the osteogenic action of FB. Methods FB were divided into 0 (control group), 0.0001,0.0010,0.1000,1.0000,10.0000 and 20.0000 mg/L groups(F~-). The levels of VEGF mRNA and protein at 48 h were measured by using RT-PCR, ELISA and immunohistochemistry (IHC) methods. Results The expression of VEGF mRNA increased obviously in group of 0.1000 mg/L(1.08 ± 0.09) in 3D FB compared with the control group(0.93 ± 0.02, all P < 0.05). Fluoride increased the content of VEGF protein obviously in groups of 0.1000,1.0000,10.0000 mg/L(0.19 ± 0.02, 0.26 ± 0.01 and 0.32 ± 0.01 ), higher than that in 2D FB culture supematant in the control group(0.14 ± 0.01, all P < 0.05) ; and in groups of 0.1000, 1.0000 rag/L(0.59 ± 0.06 and 0.52 ± 0.03) it was higher than that in 3D FB culture supematant in the control group(0.37 ± 0.05, all P< 0.01 ). The IHC results showed that the VEGF positive staining cells increased significantly in group of 0.001 mg/L (0.45 ± 0.05) in 2D FB when it was compared with control group(0.36 ± 0.03, P< 0.05); and in groups of 0.0010, 0.1000, 1.0000 rag/L(0.62 ± 0.04,0.70 ± 0.06 and 0.65 ± 0.07) are it was higher than that in 3D FB control group (0.44 ± 0.04, P < 0.05 or < 0.01 ). Conclusions The higher expression of VEGF mRNA and protein in 2D and 3D FB induced by fluoride may play an important role in stimulating the osteogenesis ability in FB.

Objective To observe the level of reduced glutathione(GSH) and oxidized glutathione(GSSG)in a mouse bone cell line MC3T3-E1 cells exposed to fluoride.Methods MTT method was used to detect cell viability of M C3T3-E1 cells exposed to varying concentrations and periods of fluoride [F-concentration:0(control),0.5,1.0,2.0,4.0,8.0,12.0,20.0 mg/L; F-periods:1,2,4 and 10 days].The Xevo TQ MS was employed to test the levels of GSH,GSSG and glutamine (Gln).Results The MC3T3-E1 cell viability was significantly higher in the 2 mg/L group(0.57 ± 0.05) 1 day after the exposure compared to the respective control(0.49 ± 0.03,P ＜0.01); conversely,cell viability was markedly lower in the 8 mg/L(0.49 ± 0.07) and 12 mg/L(0.47 ± 0.09)groups 4 days after the exposure in comparison to the control(0.63 ± 0.06,P ＜ 0.05 or P ＜ 0.01).The cell viability in the 8 mg/L group(1.52 ± 0.29) 10 days after the exposure was significantly higher than that in the control group (0.86 ± 0.23,P ＜ 0.01),however,the value in the 20.0 mg/L group (0.54 ± 0.07) was significantly lower(P ＜0.01).The level of cell GSH decreased significantly in the 20 mg/L groups 2 days[(13.92 ± 4.63)μmol/L]and 10 days [(0.53 ± 0.30)μmol/L]after exposure compared to the respective comtrols [(26.42 ± 3.67),(24.85 ± 5.68)μmol/L,all P ＜ 0.01].The level of cell GSSG markedly increased in the 2 mg/L group 2 days [(1.12 ± 0.62)μ mol/L]and the 8 mg/L group 4 days [(2.13 ± 0.62)μ mol/L]after exposure compared to the controls[(0.55 ± 0.22),(1.46 ± 0.46)μmol/L,all P ＜ 0.05].The similar change was observed in the 8 mg/L group[(2.97 ± 1.30)μmol/L] 10 days after exposure compared to the control [(1.35 ± 0.50)μmol/L,P ＜ 0.05].The level of Glndecreased significantly in the 2 mg/L group[ (62.80 ± 17.4l)μ mol/L] 4 days and in the 8 and 20 mg/L groups 10 days[ (122.26 ± 19.51), (19.38 ± 8.11)μmol/L] after exposure compared to the controls [ (83.28 ±14.32), ( 147.15± 16.95) μmol/L , all P ＜ 0.05 or P ＜ 0.01 ]. Conclusions Fluoride exposure can significantly promote the changes of GSH, GSSG and Gln levels in the osteoblast, thus affecting the intracellular redox equilibrium.

This study aims to establish a new method for quality evaluation of Gentianae Macrophyllae Radix by simultaneous determination of five iridoids (loganic acid, 6'-O-beta-D-glucopyranosylgentiopicroside, swertiamarin, gentiopicroside, sweroside), and to detect five iridoids in the root of eight species (Gentiana macrophylla, G. straminea, G. crassicaulis, G. dahurica, G. robusta, G. waltonii, G. lhassica, and G. tibetica). The separation was carried out on a Shiseido SPOLAR C18 (4.6 mm x 250 mm, 5 microm) column eluted with mobile phase of water containing 0.04% formic acid (A) and acetonitrile (B) in a gradient program. The flow rate was 0.8 mL x min(-1). The detect wavelength was set at 240 nm. The column temperature was kept at 30 degrees C. The volume of injection was 5 microL. The five iridoids were well separated with ideal linear correlations. The average recoveries were 97.35% - 106.23%. All the five iridoids were detected in the root of eight species. The contents of same species changed in a somewhat wider range. The contents in root of G. dahurica were lower than that in other species.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Gentianella ; chemistry ; Iridoid Glycosides ; analysis

ObjectiveTo investigate the expression of programmed death receptor ligand 1 ( PD-L1 ) of peripheral blood mononuclear cells (PBMCs) in patients with hepatic cystic echincccccosis (HCE) and its relation with interferon-γ.MethodsThe clinical data of 63 patients with HCE who were admitted to the First Affiliated Hospital of Xinjiang Medical University from June 2010 to February 2011 were retrospectively analyzed.All patients were divided into HCE active group (38 patients) and HCE non-active group (25 patients) according to the system established by the World Health Organization's Informal Working Group on Echinocoecosis.Twenty patients with hepatic hemangioma or healthy individuals were recruited in normal control group.The positive rate of PD-L1 expression was detected by flow cytometry and immunocytochemistry.The expression of interferon-γ was detected by enzyme-linked immtmosorbent assay (ELISA).All data were analyzed by the t test,one-way analysis of variance,LSD test and chi-square test.The relationship between the expression of interferon-γ and positive rate of PD-L1 expression was analyzed by the Pearson test.ResultsThe results of flow cytometry showed that the positive rates of PD-L1 expression in the HCE active group,HCE non-active group and normal control group were 12.1％±3.8％,10.9％ ± 2.5％ and 9.1％ ±2.5％,respectively.There was a significant difference in the positive rate of PD-L1 expression between the HCE active group and normal control group (t =3.327,P ＜ 0.05 ).The results of immunohistochemistry showed that the positive rates of PD-LI expression in the HCE active group,HCE non-active group and normal control group were 11.9％ ± 3.4％,i0.6％ ± 2.9％ and 9.5％ ± 3.6％,respectively.There was a significant difference in the positive rate of PD-L1 expression between the HCE active group and normal control group (t =2.470,P ＜ 0.05 ).The expressions of intefferon-γ in the HCE active group,HCE non-active group and normal control group were ( 141 ± 38 ) μμg/L,( 124 ± 32 ) μg/L and ( 105 ± 42 ) μg/L.There wasasignificant difference in the expression of interferon-γ between the HCE active group and normal control group ( t =3.280,P ＜ 0.05).The results of flow cytometry and immunohistochemistry revealed that the positive rate of PD-L1 expression was positively correlated with the expression of interferon-γ( r =0.59,0.61,P ＜ 0.05 ).Conclusion With the help of interferon-γ,PD-L1 may play an important role in promoting the immune.evasion of echinococcus.

A double targets of high throughput screening model for xanthine oxidase inhibitors and superoxide anion scavengers was established. In the reaction system of xanthine oxidase, WST-1 works as the probe for the ultra oxygen anion generation, and product uric acid works as xanthine oxidase activity indicator. By using SpectraMax M5 continuous spectrum enzyme sign reflectoscope reflector, the changes of these indicators' concentration were observed and the influence factors of this reaction system to establish the high throughput screening model were studied. And the model is confirmed by positive drugs. In the reaction system, the final volume of reaction system is 50 μL and the concentrations of xanthine oxidase is 4 mU x mL(-1), xanthine 250 μmol x L(-1) and WST-1 100 μmol x L(-1), separately. The Z'-factor of model for xanthine oxidase inhibitors is 0.537 4, S/N is 47.519 9; the Z'-factor of model for superoxide anion scavengers is 0.507 4, S/N is 5.388 9. This model for xanthine oxidase inhibitors and superoxide anion scavengers has more common characteristics of the good stability, the fewer reagent types and quantity, the good repeatability, and so on. And it can be widely applied in high-throughput screening research.
Enzyme Inhibitors ; pharmacology ; Free Radical Scavengers ; pharmacology ; High-Throughput Screening Assays ; Superoxides ; Uric Acid ; Xanthine ; Xanthine Oxidase ; antagonists & inhibitors