APOE) polymorphisms and age-related macular degeneration (AMD) in a Chinese population.To examine the association between apolipoprotein E  were analyzed by PCR and direct sequencing.The study consisted of 712 subjects, including 201 controls, 363 cases with early AMD, and 148 cases with exudative AMD. Genomic DNA was extracted from venous blood leukocytes. Common allelic variants of APOE ε3ε3 was the most frequent genotype, with a frequency of 72.6% in controls, 72.5% in early AMD, and 70.3% in exudative AMD. Frequency of the ε2 allele was 6.7% in controls, 7.4% in early AMD, and 8.8% in exudative AMD. Frequency of the ε4 allele was 8.7% in controls, 7.7% in early AMD, and 7.8% in exudative AMD. No statistically significant difference in APOE genotype and allele frequency distribution was observed among controls, cases with early AMD, and cases with exudative AMD. For ε2 allele carriers, the odds ratio was 1.12  for early AMD and 1.06  for exudative AMD. For ε4 allele carriers, the odds ratio was 1.04  for early AMD and 0.83  for exudative AMD.APOE polymorphisms with early or exudative AMD, suggesting that APOE is less likely to be a major AMD susceptibility gene in the Chinese population.Our data provide no evidence to support an association of  APOE), located on chromosome 19q13.2, is polymorphic, with three major allelic variants, ε2, ε3, and ε4 [APOE ε4 allele has been associated with a variety of neurodegenerative and cardiovascular diseases, most notably Alzheimer disease. Interestingly, APOE is also a ubiquitous component of drusen [APOE knockout [APOE may play a role in the pathogenesis of AMD. Several genetic association studies have shown that the APOE ε4 allele is associated with a protective effect against AMD [APOE and AMD [APOE polymorphisms with early or late AMD in an independent cohort of a Chinese population.Age-related macular degeneration (AMD) causes irreversible central vision loss and is the leading cause of legal blindness in the elderly population. Evidence suggests that AMD is a complex disease caused by actions and interactions of multiple genes and environmental factors -10. Apol, and ε4 ,15. The f drusen -21, a haknockout  or transinst AMD -28, wherinst AMD ,27-31. O and AMD -37. GiveThis case-control cohort included 201 control subjects, 363 cases with early AMD, and 148 cases with exudative AMD. Gender and age breakdown of the controls and cases were given in All subjects underwent a comprehensive ophthalmological examination, including visual acuity measurement, slit-lamp biomicroscopy, and detailed fundus examination after pupil dilation, using 0.5% tropicamide and 5% phenylephrine , performed by a retinal specialist. Smoking history was ascertained using a standard questionnaire. Stereoscopic mydriatic 30° color fundus photographs were taken from both eyes, using a digital fundus camera  centered on the fovea. Fundus photographs were assessed by two independent readers, and a retinal specialist was used as a third person to make final judgment in case of any discrepancies. Early AMD was defined as the presence of soft drusen (>63 µm) with or without pigment abnormalities . IndividNM000041.2) and 158 (R158C or c.526C>T of reference sequence NM000041.2) were then used to determine the APOE allelic variants ε2 , ε3 , and ε4 .Blood samples were collected from all participants and stored at −80 °C before DNA extraction. Genomic DNA was isolated from venous blood leukocytes using a genomic DNA extraction and purification kit  according to the manufacturer’s protocol. Briefly, lysis buffer was added to blood samples, mixed, and the tubes centrifuged. The supernatant was discarded and pellet re-suspended in a solution of proteinase K and sodium dodecyl sulfate. The mixture was incubated at 65 °C for 30 min. Genomic DNA was precipitated by isopropanol, rinsed with ethanol, and finally re-suspended in TE buffer . PCR amplification was undertaken in a GeneAmp PCR 9700 thermal cycler . A 241-bp PCR product encompassing amino acid position 112 was produced by using the forward primer 5′-GCC TCC CAC TGT GCG A-3′ and the reverse primer 5′-GGC CGA GCA TGG CCT G-3′. A 150-bp product encompassing amino acid position 158 was produced by using the forward primer 5′-ACC GAG GAG CTG CGG G-3′ and the reverse primer 5′-CTC GCG GAT GGC GCT GA-3′. PCR conditions used for amplification were 94 °C for 5 min, followed by 40 cycles of 94 °C for 30 s, 65 °C for 30 s, and 72 °C for 30 s. The final extension step at 72 °C was lengthened to 10 min. PCR products were then directly sequenced using an automatic ABI 3730XL DNA Analyzer (Applied Biosystems). Single nucleotide polymorphisms (SNPs) at amino acid position 112  and alleles  were analyzed using the χ2 test and Fisher’s exact test when the expected frequency was <5. We defined carriers of the ε2 allele to include genotypes ε2ε2 and ε2ε3 and carriers of the ε4 allele to include genotypes ε3ε4 and ε4ε4. Individuals with the ε2ε4 genotype were excluded from all analyses to avoid potential confounding effects. A logistic regression model was used to calculate the odds ratio (OR) and 95% confidence interval (CI) of early or late AMD, comparing ε2 or ε4 allele carriers to the ε3ε3 genotype as the reference, adjusted for age, gender, and cigarette smoking. A risk score model was used to evaluate the variation in APOE polymorphisms, as suggested in the literature [The data were analyzed using the terature . BrieflyA total of 712 subjects participated in the study, including 201 control subjects, 363 cases with early AMD in one or both eyes, and 148 cases with exudative AMD in at least one eye. Fluorescein angiography was undertaken in all cases with exudative AMD, and indocyanine green angiography was available in 42 cases. No atrophic AMD patient was identified among the study participants. The general characteristics of the study subjects are given in APOE genotype and allele frequency distribution among study subjects are given in APOE genotypes were under Hardy–Weinberg equilibrium in controls and cases with early or exudative AMD (p≥0.099). The ε3ε3 was the most frequent genotype with a frequency of 72.6% in controls, 72.5% in early AMD, and 70.3% in exudative AMD. Frequency of the ε2 allele was 6.7% in controls, 7.4% in early AMD, and 8.8% in exudative AMD. Frequency of the ε4 allele was 8.7% in controls, 7.7% in early AMD, and 7.8% in exudative AMD. No statistically significant differences between controls and early AMD or between controls and exudative AMD were found for any genotype or allele frequencies.The APOE alleles associated with early or exudative AMD are given in APOE risk score and early or exudative AMD was identified.The OR and risk scores of APOE ε4 or ε2 allele and any AMD was calculated under the condition of different hypothetical OR values. To detect a significantly protective effect of the ε4 allele, the sample size in this study had 83.3% power if the ε4 allele carriers had an OR of 0.5 and 30.3% power if the ε4 allele carriers had an OR of 0.7. To detect a significant risk effect of the ε2 allele, the sample size in this study had 78.0% power if the ε2 allele carriers had an OR of 2.3 and 26.8% power if the ε2 allele carriers had an OR of 1.5.The power to detect a significant association between the APOE polymorphisms and AMD has been examined in previous studies, most of which have focused on late AMD in Caucasian populations. In general, many of these previous studies suggest that the APOE ε4 allele is associated with a lower risk of late AMD [APOE and AMD, however, is not always reproducible. Several other studies reported no evidence to support a significant association between APOE and late AMD in Caucasian [APOE and early AMD was found in a large cohort of a population-based cross-sectional study in a Caucasian population [APOE and exudative AMD. In addition, no association between APOE and early AMD was found in this study. In a previous report in Japanese patients, APOE was not associated with polypoidal choroidal vasculopathy (PCV) or typical AMD [The possible relationship between late AMD -28, wherlate AMD ,27-31. Taucasian -34, Hongaucasian , and Japaucasian  populatipulation . In agrepulation , our datical AMD . In thisAPOE polymorphisms with AMD in the current study may be explained by the small or nonexistent effect of APOE on AMD in the Chinese population. It should be noted, however, that the APOE ε4 allele occurs less frequently in Chinese than in Caucasian control populations, making it more difficult to detect the effect of APOE in Chinese individuals. The frequency of the ε4 allele was reported to be 12.4%–17.9% in Caucasian control populations [APOE and AMD. We therefore cannot exclude the possibility of a weak association between APOE polymorphisms and AMD in the Chinese population.The lack of a significant association of ulations ,31,33,34APOE polymorphisms with early or exudative AMD in this study, suggesting that APOE is less likely to be a major susceptibility gene to AMD in the Chinese population. Whether there is a weak association between APOE and AMD in Chinese, however, needs to be confirmed with a much larger sample size in future studies.In summary, we provide no evidence to support a significant association of


JMHAQ, than other classes of molecules. This suggests that their corresponding log maximum fluxes through silicone from water, log JMPAQ, may be useful to extend the existing n = 63 log JMPAQ database to include more log JMPAQ values greater than 0.0. The log JMPAQ values for n = 7 phenols predicted to give log JMPAQ values greater than 0.0 based on their log JMHAQ values have been experimentally determined. These n = 7 new log JMPAQ values have been added to the existing n = 63 log JMPAQ database to give a new n = 70 database and the n = 7 literature log JMHAQ values have been added to the existing n = 48 log JMHAQ database (matched to the n = 63 log JMPAQ database) to give a new n = 55 database. The addition of the n = 7 phenols improved the correlations of these flux databases when fitted to the Roberts-Sloan equation, RS, as well as the correlation between the matched experimental (Exp.) log JMPAQ with the Exp. log JMHAQ.Phenols as a class of molecules have been reported to exhibit higher log maximum fluxes through human stratum corneum, SC, from water, log  JMPAQ, and their maximum flux through human skin in vitro from water, log JMHAQ [JMPAQ could be used to predict Exp. log JMHAQ. The rate-limiting barrier to diffusion of molecules through human skin is the stratum corneum, SC. The SC is comprised of highly dense, polar, proteinous corneocytes embedded in a lipid matrix. The lipid matrix in turn is comprised of multiple lipid bilayers containing mostly ceramides, fatty acids and cholesterol ,2,3. A tog JMHAQ . Thus exJM. Thus all the molecules are presented to the membrane at their maximum thermodynamic activity in that solvent [i.e., at their solubility limit or saturated solubility in the first few layers of the membrane, SM1. Thus, JM depends only on the solubility of the molecules in the first layers of the membrane, and unless the solvent changes the solubility limit of the membrane, the solvent/vehicle has no effect on JM. The form of the RS equation for predicting JM derives from an expansion of Fick’s law, Equation 1, so that the dependent variables are molecular weight, MW, solubility in a lipid, SLIPID, and solubility in water, SAQ. SM1 can then be estimated from the product of the partition coefficient between the vehicle  and a surrogate lipid for the membrane , (KOCT:AQ)y∙constant, and the solubility in the vehicle, SAQ: (KOCT:AQ)y∙constant∙SAQ. Expansion of that product into solubilities and taking the logs gives: log SM1 = y log SOCT − y log SAQ + log SAQ + log constant = y log SOCT + (1 − y) log SAQ + log constant.D is the diffusion coefficient of the molecule in the membrane, L is the thickness of the membrane, CM1 is the concentration of the molecule in the first few layers of the membrane and CMn is the concentration in the last few layers of the membrane which is assumed to approach zero. A linear relationship must exist between log D + log SM1 of molecules in a silicone membrane and log D + log SM1 of molecules in human skin in order for a linear relationship between log JMPAQ and log JMHAQ to exist. The flux data for molecules from which the Roberts–Sloan, RS, equation was derived is based on the application of saturated solutions (suspensions of molecules in a solvent) to the membrane being used to give maximum flux,  solvent ,7,8. Sin solvent , i.e., aJMPAQ and log JMHAQ exists is that there are only about n = 63 molecules for which log JMPAQ (output) and the necessary corresponding physicochemical properties  literature values exist which can be fitted to RS [n = 63, only 18 molecules exhibit high output values; log JMPAQ values greater than 0.0. Simple phenols present an opportunity to extend the existing n = 63 log JMPAQ database to include more log JMPAQ values greater than 0.0. The log JMHAQ values for n = 18 phenols and their corresponding physicochemical properties that are necessary to determine their fit to RS were published by Roberts, et al. in 1977 [n = 18 phenols to RS in the fit of the n = 62 edited Flynn database to RS was published in 2007 [n = 16 of the n = 62 molecules exhibited log JMHAQ values greater than 0.0 and of those n = 16, n = 11 were from among the n = 18 phenol subset [One problem with determining if the linear relationship between log ed to RS . Of thos in 1977 . The fitl subset . Thus, pn = 6 of the n = 18 simple phenol subset from Roberts, et al. [n = 63 log JMPAQ database and only n = 2 exhibit log JMPAQ greater than 0.0. In order to improve the correlation of the n = 63 log JMPAQ database with a matched n = 48 log JMHAQ database, the number of log JMPAQ and log JMHAQ greater than 0.0 in each database should be increased. Hence, n = 7 additional phenols have been selected from the n = 18 subset which exhibit physicochemical properties (input) for which RS predicts high log JMPAQ values (output). In addition, the n = 7 phenols exhibit an average log JMHAQ value significantly greater than that of the n = 48 log JMHAQ database: means ± 95% confidence intervals of 0.04 ± 0.42 log units and −1.06 ± 0.31 log units, respectively. Given the increased range and total number of entries resulting from the addition of these n = 7 compounds to the n = 48 log JMHAQ database and the n = 63 log JMPAQ database, the fit of these databases to the RS should improve, and correlation of the log JMPAQ with log JMHAQ values matched in these databases should also improve.At present, only , et al.  have been = 7 new entries to the new n = 63 log JMPAQ database, each potentially exhibiting higher log JMPAQ values than the average of the initial n = 63 log JMPAQ values, will change the relative distribution of flux values in the database, it is imperative to determine if other models would then fit the database better than they did before the addition of the n = 7 new entries. Thus, we will also determine the fit of the new databases to the Kasting–Smith–Cooper (KSC) model [Further, since the addition of the C) model  and to tC) model  and compThe phenolic compounds used are listed in The measurement of maximum flux through silicone was performed according to a literature procedure  at 32 °Cn = 3. The donor cells were sealed by Parafilm. Samples were taken from the receptor every 2–3 h after application. Following sample collection, the receptor phases were changed to ensure sink conditions, and the donor suspensions were either changed or had more solid/oil added to the existing suspension, depending upon the visible extent of depletion. After 4–5 sampling intervals, the donor suspensions were removed with methanol and the receptors were changed. The membranes were leached with methanol in the donor phase for 48–72 h with samples taken and receptor phases changed every 12–24 h to remove any residual phenol in the membrane.The donor suspensions were prepared by stirring approximately 0.5 g (1 g in the case of 3-methylphenol) of the compounds in 10 mL of water for 24 h. For all compounds, this surpassed the aqueous solubility by a factor of at least 20, which ensured saturation and excess solid/oil present in the donor phase. After the membranes were conditioned, the receptor phases were changed and the donor suspensions  were applied; To ensure that flux data was not altered by possible membrane damage, a standard solute/solvent was applied and its flux determined. A donor suspension was prepared from 400 mg of theophylline suspended with stirring in 6 mL of propylene glycol (PG) for 24 h. This suspension  was applied to all the silicone membranes after they were leached with methanol (see above). Samples were taken from the receptor every 24 h after application for at least 72 h so that at least 3 samples were obtained. Following sample collection, the receptor phases were changed and the donor suspensions were changed every other sampling interval. After 3–5 sampling intervals, the diffusion cells were disassembled and the membranes were placed in a methanol bath for maintenance leaching.ε) and molar absorptivities (ε) used for the phenolic compounds are listed in JMPPG, for each membrane was found to be within the standard deviation of the literature value of −2.68 ± 0.12 log units [The flux values of the first and second application were determined by UV absorption. The wavelengths (λog units .JMAQ:Nonlinear regression was performed by SPSS 20.0 (Rel. 20.0.0). The compounds were fitted to the Roberts–Sloan equation for maximum flux, log JMPAQ greater than 0.0, and even it was very close. As a subset the n = 7 simple phenols had an average log JMPAQ significantly greater than the average log JMPAQ of the n = 63 log JMPAQ database: means ± 95% confidence intervals 1.03 ± 0.45 log units and −0.42 ± 0.29 log units, respectively. The average log JMPAQ in the n = 70 log JMPAQ database has not significantly increased, but is no longer significantly less than 0.0: mean ± 95% confidence interval −0.27 ± 0.29. Unfortunately, the addition of the n = 7 phenols did not significantly increase the average log JMHAQ of the n = 55 log JMHAQ database relative to the n = 48 log JMHAQ database: means ± 95% confidence intervals, −0.918 ± 0.29 log units and −1.058 ± 0.31 log units, respectively.The results are displayed in n = 7 phenols to the n = 63 log JMPAQ database and the n = 48 log JMHAQ database improved the fit of these databases to the RS as expected. The fit of the new n = 70 log JMPAQ database gave an r2 of 0.907, an average absolute residual log JMPAQ (Δlog JMPAQ) of 0.300 log units and the coefficients x = −1.606, y = 0.695 and z = 0.00490 were all significant (p < 0.05):The addition of these n = 70 log JMPAQ database is an improvement over the n = 63 log JMPAQ database, which had r2 = 0.896 and Δlog JMPAQ = 0.310 log units, but had similar coefficient values: x = −1.607, y = 0.701, z = 0.00492. The fit of the new n = 55 log JMHAQ database gave an r2 of 0.883, an average absolute residual log JMHAQ (Δlog JMHAQ) of 0.282 log units and the coefficients x = −3.005 and y = 0.654 were significant (p < 0.05), but the coefficient z = 0.00112 was not significant (p = 0.25):The fit of the z coefficient indicates a need to further extend the n = 55 log JMHAQ database, since the significance of MW to maximum flux is well-established [n = 55 log JMHAQ database to the RS is an improvement over the n = 48 log JMHAQ database, which had r2 = 0.867 and Δlog JMHAQ = 0.331 log units and the coefficients x = −2.763, y = 0.635 and z = 0.00207. The x and y coefficients for the new log JMHAQ database are substantially closer to those coefficients determined for the n = 62 edited Flynn log JMHAQ database: x = −3.008, y = 0.732, z = 0.0048. Relevant results are displayed in JMPAQ versus log JMPAQ calculated  from the coefficients for the fit of the n = 70 database to RS, and JMHAQ versus log JMHAQ Calc. from the coefficients for the fit of the n = 55 database to RS. The lack of statistical significance for the ablished . The fitSOCT, log SAQ and MW, against log JMPAQ (n = 70) and against log JMHAQ (n = 55) gave the following r2 values: (a) log JMPAQ versus log SOCT, r2 = 0.677; versus log SAQ, r2 = 0.554; versus MW, r2 = 0.541; (b) log JMHAQ versus log SOCT, r2 = 0.603; versus log SAQ, r2 = 0.526; versus MW, r2 = 0.520. All regression equations had statistically significant (p < 0.05) slope and intercept estimates. In each case, the best regression of the individual independent variables against flux values was by log SOCT. It is worth noting that the n = 7 phenol subset gives the following r2 values and significance profiles when the individual independent variables log SOCT and log SAQ are plotted against log JMPAQ and log JMHAQ: (a) log JMPAQ versus log SOCT, r2 = 0.180, without a statistically significant slope (p = 0.34) or intercept (p = 0.51); versus log SAQ, r2 = 0.949, with a statistically significant slope (p < 0.05), but without a statistically significant intercept (p = 0.50); (b) log JMHAQ versus log SOCT, r2 = 0.335, without a statistically significant slope (p = 0.17) or intercept (p = 0.18); versus log SAQ, r2 = 0.933, with statistically significant (p < 0.05) slope and intercept. The improved dependence of maximum flux from water on the aqueous solubility of highly water-soluble compounds is demonstrated here as a property of both silicone and human stratum corneum, and will be a topic of future investigations. When two individual independent variables, log SOCT and MW, from the n = 70 log JMPAQ database were fitted to the KSC equation (Equation 3) the following x, y and z coefficients to the parameters were obtained along with r2 and Δlog JMPAQ values: x = −0.923, y = 0.794, z = 0.0089, r2 = 0.797 and Δlog JMPAQ = 0.431. All but the y coefficient (p = 0.069) were statistically significant (p < 0.05):Plots of the individual independent variables, log JMPAQ versus log JMPAQ values calculated from the coefficients for the fit of the n = 70 database to Equation 7. Although the r2 was substantially improved by including MW with log SOCT as independent variables in the regression against flux, the r2 was poorer than the r2 for the fit of all three independent variables to the Roberts–Sloan Equation (Equation 5). Similarly, when two individual independent variables, log SOCT and MW, from the n = 55 log JMHAQ database were fit to the KSC equation (Equation 8) the following x, y and z coefficients to the parameters were obtained along with r2 and the Δlog JMHAQ values: x = −1.252, y = 0.602, z = 0.0080, r2 = 0.723 and Δlog JMHAQ = 0.441. The estimates for the coefficients were all statistically significant (p < 0.05): JMHAQ versus log JMHAQ values calculated from the coefficients for the fit of the n = 55 database to Equation 8. Again, although a substantial improvement in r2 was obtained by including MW with log SOCT as independent variables in the regression against flux, the r2 was poorer than the r2 for the fit of all three independent variables to the RS equation (Equation 6). The fit of both databases to the MACR equation (Equation 4), which is the remaining model used to predict maximum flux, is simply the regression of MW against log JMPAQ or log JMHAQ shown above to give a somewhat poorer fit than regression of the two individual independent variables, log SOCT or log SAQ, against log JMPAQ or log JMHAQ. Finally, it should be noted the popular Potts–Guy Equation [Equation  was not n = 52 database of compounds contributing to both the n = 70 log JMPAQ database and the n = 55 log JMHAQ database also gives a higher correlation between Exp. log JMPAQ and Exp. log JMHAQ than the previous n = 45 database. A linear regression yielded the expression Exp. log JMHAQ = 0.859 Exp. log JMPAQ − 0.837, r2 = 0.856, which is an improvement over r2 = 0.838 for the n = 45 database. JMPAQ versus Exp. log JMHAQ for n = 52. The new n = 7 phenols improved all aspects of the log JMPAQ database with regards to comparison with the matched log JMHAQ database. Along with strengthening the validity of the assertion that silicone membranes are good surrogates for human stratum corneum, these improvements demonstrate that this surrogate nature holds for a wider range of log JMPAQ and log JMHAQ values than had been previously reported. The addition of the


Flemingia macrophylla root (WEFM) were investigated. The results showed that WEFM exhibited radical scavenging and reducing activities, as well as ferrous ion chelating property. In addition, WEFM also protected phospholipids against oxidation, indicating that WEFM could protect biomolecules from oxidative damage. Meanwhile, in the range of 50–100 μg/mL, the tyrosinase inhibitory activity of WEFM increased with an increase in sample concentration and was superior to that of the water extract of Glycine tomentella root (WEGT). A high performance liquid chromatography analysis was used to determine the phenolic components, revealing that daidzin, daidzein, genistin, and genistein were present in WEFM and WEGT. Acting as an antioxidant and a tyrosinase inhibitor, these bioactive constituents could contribute to the protective effects of WEFM. Overall, the results showed that WEFM might serve as a natural antioxidant and tyrosinase inhibitor.The antioxidant and antityrosinase activities of the water extract of  They are also known to contaminate the food with harmful substances. At the same time, lipid oxidation induced by reactive oxygen species (ROS) may lead to cellular damage and promote the pathological progression of atherosclerosis, carcinogenesis, and diabetes . Therefoin foods . In addiin foods . Therefoin foods . In addiin foods , 6 as wein foods , 8.Flemingia macrophylla (FM) or Glycine tomentella (GT) is available in teabags on the shelves of supermarkets in Taiwan Island and the south of China. The roots of FM and GT, which were misused or substitutionally used on the market, have long been used to treat rheumatism, arthritis and muscle pains, and so forth [Phytochemical-rich natural products, which have a complex array of antioxidants, could exhibit protective effects against oxidative stress in different models. For example, herbal teas are one of the oldest beverages in Southeast Asia, such as China, Taiwan, Vietnam, and Philippines. Recently, a herbal tea prepared from the roots of so forth . Both coso forth , 11. Theso forth –14, therFlemingia macrophylla and Glycine tomentella were obtained from local Chinese herbal medicine stores in Tainan, Taiwan.1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulphonate) diammonium salts (ABTSs), lecithin, potassium ferricyanide, ferrozine, thiobarbituric acid (TBA), and mushroom tyrosinase were purchased from Sigma . Daidzin, daidzein, genistin, and genistein were purchased from extrasynthese . Acetonitrile was of the LC grade . The roots of Flemingia macrophylla and Glycine tomentella were named as WEFM and WEGT, respectively. The yield obtained was 23.8% (11.9 g) and 16.6% (8.3 g), respectively.All dried samples were cut into small pieces and ground into fine powder. Each sample (50 g) was extracted with boiling water (1000 mL) for 30 min, and the filtrate was evaporated to dryness under vacuum. The final dried extracts of roots of μm filter. The injection volume was 20 μL, and the flow rate was 0.8 mL/min. The separation temperature was 25°C. The column was a Cosmosil 5C18-AR . The method involved the use of a binary gradient with mobile phases containing (A) phosphoric acid in water  and (B) H2O/CH3CN: 20/80 (v/v). The solvent gradient elution program was as follows: 0–5 min, 100–90% A, 0–10% B; 5–20 min, 90–85% A, 10–15% B; 20–25 min, 85–80% A, 15–20% B; 25–30 min, 80–60% A, 20–40% B; 30–40 min, 60–40% A, 40–60% B; 40–50 min, 40–10% A, 60–90% B; 50–60 min, 10–0% A, 90–100% B; finally, 60–70 min, 0% A, 100% B. Based on the plot of the peak-area ratio (y) versus concentrations , the regression equations of daidzin, daidzein, genistin, and genistein and their correlation coefficients (r) were as follows: daidzin, y = 0.0234x + 0.0012 (r = 0.9969); daidzein, y = 0.0533x − 0.0018 (r = 0.9970); genistin, y = 0.0356x − 0.0002 (r = 0.9969); genistein, y = 0.0469x + 0.0029 (r = 0.9972).A high-performance liquid chromatography (HPLC) was performed with a Hitachi Liquid Chromatography System , consisting of two model L-7100 pumps and a model L-7455 photodiode array detector. Samples (10 mg/mL) were filtered through a 0.45 w/v)) individually to a 10 mL volumetric flask. After 5 min, 0.1 mL Folin-Ciocalteu reagent (50% (v/v)) was added, and the volumes were made up to 10 mL with H2O. After incubation at 30°C for 1 h, the absorbance at 750 nm was measured and compared to the gallic acid calibration curve.Total polyphenols were determined as gallic acid equivalents . DiffereDifferent concentrations of the samples were added with 0.1 mL (2-aminoethyl) diphenyl borate (0.2% in ethanol) individually. After 20 min of incubation, the absorbance at 405 nm was measured and compared to a rutin calibration curve .The assay was carried out as previously described . Each sa•+ as previously described [•+ was generated by reacting 1 mM ABTS with 0.5 mM hydrogen peroxide and 10 units/mL horseradish peroxidase in the dark at 30°C for 2 h. After 1 mL of ABTS•+ was added to samples, the absorbance at 734 nm was recorded after 10 min.This assay determined the capacity of samples to scavenge ABTSescribed . The ABTThe reducing power of sample was determined as previously described . Potassi2+ was measured as previously described [2  and ferrozine  for 10 min, and the spectrophotometric absorbance was determined at 562 nm. A lower level of absorbance indicated a stronger chelating activity.The chelating activity of samples on Feescribed . Each sa3, ascorbic acid, and samples (0.2 mL) were mixed to produce a final concentration of 3.12 μM FeCl3 and 125 μM of ascorbic acid. The mixture was incubated for 1 h at 37°C by the thiobarbituric acid (TBA) method [Lecithin (500 mg) was sonicated in an ultrasonic cleaner  in phosphate buffer  for 2 h in an ice-cold water bath. The sonicated solution, FeClThe mushroom tyrosinase was used for the bioassay. The inhibition on tyrosinase-catalyzed oxidation of L-DOPA was determined as previously described . The reaP < 0.05.Statistical analysis adopted in this study involved the use of the Stat View statistical package (SAS institute Inc.). Analysis of variance was performed by ANOVA procedures. Significant differences between means were determined by Duncan's multiple range tests at a level of Previous studies have indicated that the free radical scavenging activities observed in FM and GT may be due to various natural polyphenols such as daidzin, daidzein, genistin, and genistein which displayed antioxidative effects ; thus, d50 value of WEFM and WEGT was 65.28 μg/mL and 298.66 μg/mL, respectively. The ABTS cation radical inhibition assay, which has been demonstrated to evaluate the total antioxidant capacity of natural products, was also determined. In this study, the water-soluble counterpart of WEFM and WEGT exhibited different scavenging activity against ABTS radical cation in a concentration-dependent manner. 50 value of WEFM and WEGT was 23.05 and 146.23 μg/mL, respectively. These results indicate that the bioactive constituents of the WEFM were potential candidates in scavenging harmful radicals. As shown in μg/mL exhibited a dose-dependent protective effect, 46.7–67.5%, on the liposome damage induced by the Fe3+/H2O2 reaction. The lipid oxidation inhibition of WEFM at the concentration of 50 μg/mL was about 1.9-fold greater than that of WEGT. Similarly, μg/mL possessed protective effects against liposome oxidation. In particular, genistein showed the highest level of protection among the four reference constituents.Liposome protection was used as an index to assay the protective effects of the WEFM and WEGT on lipid oxidation. As shown in μg/mL were 13.7, 13.4, 6.3, and 6.9 μg VitC equivalents/mL. Genistein and genistin had stronger reducing activities than daidzein and daidzin. It was observed that the total phenolics of WEFM had a good correlation with its liposome protection activity and reducing power. This implied that the total polyphenols of WEFM could contribute to the liposome protection and reducing activity. Finally, WEFM did show the protective capacity against lipid molecule oxidation. On the other hand, it has been suggested that an antioxidant capacity is due to the development of a reducing ability when reactiing with free radicals and terminating the radical chain reaction . Accordiμg/mL, WEFM and WEGT exhibited 26.3–45.8% and 11.7–18.9% inhibitory effects on tyrosinase activity, respectively. The tyrosinase inhibition of WEFM at the concentration of 100 μg/mL was about 2.4-fold greater than that of WEGT. In this test, the tyrosinase inhibitory effects of WEFM increased with an increase in sample concentration and were superior to that of WEGT. Similarly, μg/mL. Previous reports have suggested that metal chelators could be used as tyrosinase inhibitors [μg/mL, respectively. This result showed that WEFM exhibited a greater inhibitory effect on chelating activity than that of WEGT. The chelating effects of genistein, genistin, daidzein, and daidzin at 100 μg/mL were 42.6, 48.8, 77.8, and 75.8%, respectively. Genistein and genistin had weaker chelating activity than daidzein and daidzin. However, daidzin, with the tyrosinase inhibition effect of 35.8% at 100 μg/mL, showed the lowest level of inhibition among the four reference constituents.In addition, the tyrosinase inhibitory activities of WEFM and WEGT are compared in hibitors ; thus, tThe destructive stress derived from reactive oxygen species will induce oxidation in biomolecules  and aggravate the progression of cells to a pathological status . On the Free radicals can induce pathological events such as inflammation, aging, and carcinogenesis . In thisWhen lipid peroxidation occurs in cell membranes, it releases arachidonic acid, which is responsible for long-term damage to cells. Further, decreasing of low-density lipoprotein oxidation by natural antioxidants would contribute to the prevention of atherosclerosis . As showin vitro. This finding suggests that WEFM decreases the level of stress resulting from a reduction in glutathione and the quinone-induced cell damage. Previous studies have supported the role of metal ion chelation in the process of tyrosinase inhibition, and a linear correlation existed between chelating activity and tyrosinase inhibition [It has been reported that tyrosinase, also known as a polyphenol oxidase, plays a critical role in catalyzing the melanogenesis and promoting the reactive metabolites produced in the process of melanin formation . Meanwhihibition . PolypheFlemingia macrophylla is interesting as herbal antioxidant for pharmaceutical products and cosmetics.Our experimental study revealed that WEFM exerted significant antioxidative effects and antityrosinase activity in the acellular systems of this study. Therefore, the root extract of


The objective of this study was to evaluate changes in left ventricular ejection fraction (LVEF) and left ventricular remodeling after coronary microembolization (CME) and to investigate the protective effects of methylprednisolone (MTP).n = 9) received 120,000 microspheres and Group 2 (n = 7) received 120,000 microspheres following intravenous administration of 30 mg/kg MTP. Contrast-enhanced magnetic resonance imaging (CeMRI) was performed at baseline, 6 h after intervention, and 1 week later.CME was induced by injection of microspheres (42 μm Dynospheres) into left anterior descending artery of mini swine. The animals were divided into two groups. Group 1 (P >0.05). Hyperenchancement at the anterior wall of the left ventricle was shown by MRI at 6 h in Group 1 but not in Group 2. The hyperenhanced area in Group 1 was 7.77 ± 1.49% of left ventricular mass.In Group 1, LVEF was significantly decreased at 6 h but recovered 1 week. This was accompanied by continuing left ventricular remodeling. In Group 2, LVEF remained unchanged at all assessment times. LVEF measured at 6 h and 1 week after CME in Group 1 and Group 2 was 0.39 ± 0.06 and 0.44 ± 0.04, and 0.44 ± 0.04 and 0.48 ± 0.03, respectively (Both The consequence of CME is left ventricular dilation with preserved LVEF. Pretreatment with MTP appears to have a cardioprotective effect on left ventricular remodeling. Coronary microembolization (CME) can potentially lead to a perfusion-contraction mismatch, inflammation, and regional myocardial contractile dysfunction in the absence of coronary artery stenosis-7. It isGlucocorticoids have been used in various diseases to prevent or minimize inflammation, but conSixteen mini swine of either sex (20 to 25 kg body weight) were sedated with intramuscular ketamine (5 to 10 mg/kg) and diazepam (5 to 10 mg/kg). Anesthesia was maintained with 3% pentobarbital sodium intravenously (3 mg/kg/h) until the experiment was completed.n = 7) received 30 mg/kg MTP (in 10 mL 0.9% NaCl solution) injected intravenously 30 min before preparation for surgery. Animals in Group 1 (n = 9) received no MTP.The right femoral artery and vein were dissected and 7 F and 6 F vascular sheaths were placed in each vessel, respectively. Prior to coronary cannulation, animals were anticoagulated with intravenous heparin . For coronary angiography, a 6 F XB 3.5 guiding catheter was used for the left coronary artery system via the femoral approach. A 2.8 F infusion catheter  was then placed in the left anterior descending (LAD) artery between the second and third diagonal branches. Microembolization was induced by continuous injection of 120,000 microspheres  into the LAD followed by a 6 mL saline flush. Animals in Group 2 . Transverse, two-chamber, and four-chamber left ventricular long-axis scout images were obtained to determine the final short-axis image plane. Cine MR images were acquired in contiguous short-axis planes from the apex to the base of the heart to measure left ventricular function. After the cine MR images were obtained, the animals received an intravenous bolus of 0.20 mmol/kg Gd-DTPA  at a rate of 4 mL/s by means of an infusion pump. This was followed by a 10 mL flush of saline at a rate of 4 mL/s. After 10 min, delayed images were acquired using a Turbo_flash_T1WI_PSIR_segmented sequence. The inversion time was determined at 10 min with TI-Scout sequence. Results were analyzed at Siemens Syngo Leonardo workstation.t test were used to compare LVEF, LVEDV, and LVESV between groups. Values of P <0.05 were considered statistically significant.Statistical analysis was undertaken using SPSS version 11.0 software. Data were presented as means and standard deviations (±SD). Within group changes in hemodynamics (left ventricular ejection fraction (LVEF), left ventricular end diastolic volume (LVEDV), and left ventricular end systolic volume (LVESV)) were compared using a general linear model of variance for repeated measures. Independent Coronary thrombolysis in myocardial infarction (TIMI) grade 3 coronary flow was maintained immediately after microembolization and hemodynamic parameters remained stable before and after CME.P <0.01) and returned to baseline 1 week after CME. In addition LVEDV and LVESV were higher 1 week than at baseline in this group (P <0.01). In Group 2, there was no significant change in LVEF, LVEDV, and LVESV before or after CME. There was no difference in LVEF, LVEDV, or LVESV at 6 h between Group 1 and Group 2 (P = 0.08 for LVEF). However, at 1 week before and after changes in LVEDV and LVESV were significantly lower in Group 2 than in Group 1 (P <0.01). Before and after changes in LVEF at 6 h were significantly lower in Group 2 than in Group 1 (P <0.01). The results are shown in Table In Group 1, LVEF decreased at 6 h . It is a marker for fatal events, especially in patients undergoing percutaneous coronary intervention (PCI)-19. RuptIn the present study, we addressed changes in left ventricular function after CME and investigated the possible protective role of methylprednisolone. Our objective was to better understand the development of CME and to evaluate the possible benefit of glucocorticoid pretreatment. Using an experimental model developed by our team, we demonstrated impairment of left ventricular function (evidenced by changes in LVEF) at 6 h, recovers spontaneously 1 week later after CME, but left ventricular remodeling continues. We alsoBased on these findings, the observed changes were thought to be related to an inflammatory response rather than to micro-infarction after CME. Previous studies have demonstrated that myocardial infarction following CME affects only about 2% to 5% of the perfusion territory, and that apoptosis is almost negligible,5. Our rPrevious studies have shown that inflammatory responses induced by microinfarction, is mediated by monocytes, macrophage infiltration, and the release of inflammatory factors,8. In a In our experiments LVEF was preserved 1 week after CME due to left ventricular remodeling, which occurred as a consequence of the persistent inflammatory response in the embolized zone. A previous study, using MRI to evaluate changes after CME, demonstrated a significant decline in LVEF 1 h and 1 week after intervention. The difIn our previous study, hyperenchancement was observed 6 h after CME. This hyAn important finding of the present study was that pretreatment with MTP eliminated the hyperenhancement at 6 h after CME and protected the decline of LVEF as well as left ventricular remodeling. The anti-inflammatory effects of glucocorticoids have been widely used for the treatment of a variety of diseases in clinical practice. In experimental studies, glucocorticoids have been shown to attenuate the interaction of leukocytes with endothelium and to iA previous study reported that MTP was able to protect against progressive myocardial contractile dysfunction after CME, suggested that it may have a cardioprotective effect in this setting. These fAs there was no control group in our study it was not possible to eliminate changes due to the surgical procedure. However, previous studies have demonstrated this type of animal model closely resembles CME seen in clinical practice and in previous studies control groups were not used. A second limitation is that this animal model resembles but does not exactly replicate the sequence of events that occurs following complications of percutaneous coronary intervention. In clinical practice the thromboemboli are frequently much larger, are of non-uniform diameter, and do not always result in myocardial damage. In future studies, a longer observation period should be adopted to observe the long-term effects of CME and glucorcoticoids on left ventricular remodeling.Our results demonstrate that experimentally induced CME resulted in left ventricular dilation with preserved LVEF. Pretreatment with MTP had a cardioprotective effect on left ventricular remodeling. These findings are of potential significance for the clinical management of CME.ACS: Acute coronary syndrome; CeMRI: Contrast enhanced magnetic resonance imaging; CME: Coronary microembolization; LAD: Left anterior descending; LVEDV: Left ventricular end diastolic volume; LVEF: Left ventricular ejection fraction; LVESV: Left ventricular end systolic volume; MTP: Methylprednisolone; PCI: Percutaneous coronary intervention; TIMI: Thrombolysis in myocardial infarction.The authors have no competing interests.JM carried out the animal studies, performed the statistical analysis, and drafted the manuscript. JQ participated in the design of the study and helped to draft the manuscript. SC participated in the animal model preparation and construction. ZC participated in the animal model construction. HJ participated in the MRI and data collection. MZ participated in the MRI. YZ participated in the design of the study and helped to perform the statistical analysis. JG conceived of the study, and participated in its design and helped to draft the manuscript. All authors read and approved the final manuscript.Jianying Ma: PhD, MD, Associate Professor of Medicine/Cardiology, Department of Cardiology, Zhongshan hospital, Fudan University; Shanghai institute of Cardiovascular disease, Shanghai, China. Juying Qian: MD, FSCAI, Professor of Medicine/Cardiology, Deputy Director of Department of Cardiology, Zhongshan hospital, Fudan University; Shanghai institute of Cardiovascular disease, Shanghai, China. Shufu Chang: MD, Attending physician, Department of Cardiology, Zhongshan hospital, Fudan University; Shanghai institute of Cardiovascular disease, Shanghai, China. Zhangwei Chen: MD, Attending physician, Department of Cardiology, Zhongshan hospital, Fudan University; Shanghai institute of Cardiovascular disease, Shanghai, China, Shanghai, China. Hang Jin: PhD, MD, Associate professor of Radiology, Department of Radiology, Zhongshan hospital, Fudan University, Shanghai, China. Mengsu Zeng: PhD, MD, Professor of Radiology, Director of Department of Radiology, Zhongshan hospital, Fudan University, Shanghai, China. Yunzeng Zou: Professor of Institute of Biomedical Science; Professor of Medicine/Cardiology, Department of Cardiology, Zhongshan hospital, Fudan University; Shanghai institute of Cardiovascular disease, Shanghai, China. GE Junbo, MD, FESC, FACC, FSCAI, Professor of Medicine/Cardiology, Director of Department of Cardiology, Zhongshan Hospital, Fudan University; Director of Shanghai Institute of Cardiovascular Diseases, Shanghai, China.


Diagnostic problems in clinical trials are sometimes ordinal. For example, colon tumor staging was performed according to the TNM classification. However, clinical data are limited by markedly small sample sizes in some stage.We propose a distribution-free test for detecting ordered alternatives in a completely randomized design. The new statistic is based on summing all correctly (ascending) ordered samples.The exact mean and variance of the null distribution are derived and it is shown that this distribution is asymptotically normal. Furthermore, we show using Monte Carlo simulation that the proposed test is a significant improvement over the Terpstra-Magel test. That is, power is decreased where the investigator falsely assumes an a priori ordering relationship.We conclude that these tests frequently detect an ordered trend when, in fact, one does not exist. However, the new test can reduce the error rate, at least not to the extent in which the Jonckheere-Terpstra test does. The hypothesis to be tested is H0 : F1(x) = F2(x) = ⋯ = Fk(x) for all x and H1 : F1(x) ≥ F2(x) ≥ ⋯ ≥ Fk(x), for all x with F1(x) > Fk(x) for some x, where F1(x), F2(x), ⋯, Fk(x) are continuous distribution functions.This paper focuses on considering nonparametric tests for the non-decreasing ordered alternative of Fi(x) = F(x - μ - θi), where μ is a location parameter and θi represents the effect of group i, i = 1, 2, …, k. This implies that the underlying populations may differ only in location. Throughout the article, let k populations with distribution functions Fi (x), i = 1, 2, …, k, respectively.In this article, we assume the location model with Nonparametric order restricted inference has been extensively investigated in past literature and new studies are continuing to emerge. For instance, Puri , Puri ank (≥ 3)-sample distribution-free statistic for ordered alternatives was proposed by Jonckheere , which we write as,Let us now consider k ties for i ≠ j. it is straightforward to show Ru denotes the rank of xu with respect to x1, x2,…, xk, Rv denotes the rank of xv with respect to x1, x2,…, x2k-3, u < v-k, Ru< Rv, and X1, X2, X3 denote the tied observations then the covariance term has the form COV  where, Ru denotes the rank of xu with respect to Iu = I(Ru = u), and Rv denotes the rank of xv with respect to Iv = I(Rv = v).Consider first the case of H0, . Next, consider E [IuIv]. This expectation contains 2k-3 observations, so that under H0, each of the (2k-3)! permutations of the observations are equally likely. However, there are only the numbers of {1 : Ru-1}∩{1 : Rv-1} possible ways, say ISS, plus the numbers of {Ru+1 : 2k-3}∩{1 : Rv-1} possible ways, say ILS, plus the numbers of {Ru+1 : 2k-3}∩{Rv+1 : 2k-3} possible ways, say ILL, to preserve X1, X2, and X3. Furthermore, there are Xu, Xv, Xu, and Xv. Hence, these arguments imply that,Under t1 + t2 + t3 = 3, t1, t2, and t3 = 0, 1, 2, 3.where l1, l2, and l3, there are Ru, Rv and all possible treatment locations (m and n), in the case of one tie X1, (7) reduces to,Now, for a given t1 + t2 + t3 = 1, t1, t2, and t3 = 0, 1.where V[TN] - V[PN] = o(1). Asymptotic normality results are attainable.From (6) and (8) it follows that The institutional review board of Chang Gung Memorial Hospital approved the present study. Detailed information about patients with colon cancer, such as patient- and tumor-related factors and follow-up status, was retrieved from the Colorectal Section Tumor Registry at Chang Gung Memorial Hospital, Taiwan. All the data in this registry were prospectively collected.Between January 2006 and December 2010, 154 consecutive patients with histologically confirmed colonic adenocarcinoma underwent curative surgeries at the Chang Gung Memorial Hospital in Chiayi. The stage IV colon cancer, non-curative surgeries, rectal cancer and mucinous adenocarcinomawere excluded in this study. Tumor staging was performed according to the TNM classification described in the 6th edition of the cancer staging manual of the American Joint Committee on Cancer . The different tumor staging require a different treatment to optimize patient and hospital outcomes. An ordinal logistic regression model was developed with predictors as follows: age, gender, tumor location, histologic differentiation, preoperative albumin level, preoperative carcinoembryonic antigen level, and underlying medical illnesses.k).To illustrate the KTMB test, assume an outcome with four stages and a set of cases consisting of one case from each stage. The case from Stage I has risks of 0.50, 0.25, 0.15 and 0.10 for Stage I, II, IIIA and IIIB, respectively. The case from Stage II has risks 0.26, 0.52, 0.17 and 0.05; the case from Stage IIIA has risks 0.06, 0.32, 0.42 and 0.20; the case from Stage IIIB has risks 0.12, 0.18, 0.30 and 0.40. The risk for Stage IIIB  is higher for the case that belongs to this stage (0.40) than for the other cases . The risk for event is second-highest for the case from Stage IIIA . However, the risk for event is lowest for the case from Stage II . The risk for event is third-highest for the case from Stage I . Therefore, the risks correctly identify the cases from Stage IIIA and IIIB but not Stage I and II, resulting in a score of 2 for this set (H0 : FI(x) = FII(x) = FIIIA(x) = FIIIB(x) for all x and H1 : FI(x) ≥ FII(x) ≥ FIIIA(x) ≥ FIIIB(x), where FI(x) ≠ FIIIB(x) for some x.Hence, the set of hypotheses was Five test statistics and the corresponding p-values are given in Table Xij as: Xij = θi + ϵij, where ϵij is the iid log-F distribution, and θi are location parameters.To determine if the underlying population came from different skew and kurtosis distributions that impact on the power of the test statistic, we used log-F distributions with combinations of 2, 4.5 and 10 degrees of freedom to generate the random variable. We can therefore define random variable ni) and location parameters (θi) we examine the different combinations of k = 3 and 4, ni = 4, 5, 8 and 10, θi = 0, 0.25, 0.5, 0.75, 1 and 1.25. We investigated designs under assumed alternatives which are of the forms of concave and convex. Programs to compare powers were written in R 2.9.2 . The estimations were conducted by simulating 10,000 different sets of samples. Furthermore, we estimated the power by counting the number of times H0 was rejected and using the value to divide by 10,000. Ideally, we believe that the test should have higher power than a general alternative test when H1 is true, and should have low power for any alternative that does not fit the profile given in H1.For the numbers of treatment (k), sample sizes /TM, ranges from -6.29% to 11.27% with the average gain percentage in power being 2.63% (difference of percentage).k = 3. The DP ranges from -13.2% to 27.68% with the average loss percentage in power being 8.83% for k = 4.Consider the corresponding alternatives of the form of concave and convex shapes. The powers of the KTMB test outperforms (lower power) the KTP, JT, MJT, and TM tests when balanced design. The loss percentage in power, DP = /KTMB, ranges from -7.28% to 20.00% with the average loss percentage in power being 4.25% for When the sample sizes corresponding to the non-decreased trend location parameters are comparatively large, the KTMB test is better than KTP, MJT, JT, and TM tests. The DP ranges from -7.29% to 9.30% with the average loss percentage in power being 2.47% for k = 3. The DP ranges from -6.52% to 30.93% with the average loss percentage in power being 12.07% for k = 4. However, the KTP test slightly better than KTMB test when the underlying population is skewed to the right  . They meThe authors declare that they have no competing interest.CHC designed the study, prepared the manuscript and performed statistical analyses. WY drafted and assisted with the revision of the article. CCC and YYH participated in and carried out the field work. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2288/13/148/prepubAlgorithm for computing .Click here for file


Background. Increased insulin resistance is an extragastrointestinal manifestation of Helicobacter pylori (HP) infection. HP changes the level of inflammatory markers and cytokines and changes the adipocyte function by altering the adiponectin level. Given the high prevalence of HP and diabetes in our society, we evaluated the association between HP and serum adiponectin level. In this cross-sectional study, 211 diabetic patients under treatment other than insulin were studied. These patients were divided into two groups of HP+ and HP− based on their HP IgG antibody serology and their blood adiponectin levels were measured. Data was analyzed using independent t-test, Chi-square test, and Fisher's exact test. Results. Seventy-two patients with an average age of 51.56 ± 8.34 years were HP− and 139 patients with an average age of 50.35 ± 9.01 years were HP+. The mean serum adiponectin level in HP− and HP+ groups was 4.54 ± 5.43 and 5.64 ± 3.88 ng/mL, respectively. Insulin resistance degree was significantly higher in HP+ group  but no significant difference was found between the mean serum adiponectin level in HP− and HP+ groups (P = 0.140). Conclusions. Although the insulin resistance degree was significantly higher in HP+ diabetic patients, no significant relationship was found between HP infection and serum levels of adiponectin. Insulin resistance (IR) can be considered as a clinical state of low insulin sensitivity at one or more of the sites involving glucose metabolism in which a normal or elevated insulin level will cause an attenuated biologic response like insulin-stimulated glucose disposal , 2. It iHelicobacter pylori (HP) is a human pathogen that infects the gastric mucosa and causes inflammatory process that increases in chronic gastritis, peptic ulcer, gastric lymphoma of mucosa-associated lymphoid tissue, and adenocarcinoma [arcinoma . It is aarcinoma . The HP arcinoma .α in diabetic patients [Based on some researches, the prevalence of HP infection in diabetics is higher than nondiabetics and its prevalence is associated with duration of diabetes , 9. Alsopatients .These inflammatory factors may cause metabolic changes in these patients which may lead to insulin resistance. Different studies evaluated the association of HP with insulin resistance some of which found a positive association –14 whileOn the other hand, adiponectin is a protein hormone released from adipose tissue and adipocytes. Low adiponectin level is associated with complications such as increased insulin resistance, diabetes, and metabolic syndrome .Based on studies, one major cause of adiponectin level change is adipocyte changes due to inflammatory cytokines. Among them, interleukin-1 and CRP have a special place. HP causes change in these markers and cytokines due to cagA gene and, theoretically, it can change the adiponectin level by changing the adipocyte function and adiponectin level change will cause insulin resistance and eventually diabetes .Given the high prevalence of HP infection and its role in insulin resistance, the aim of this study was to evaluate the association between serum adiponectin level and HP infection.This cross-sectional study was carried out from May to December 2012 on diabetic patients referred to diabetes clinic of Shahid Beheshti Hospital of Qom. In case of receiving insulin, smoking, pregnancy, and a history of HP treatment , patients were excluded from the study. Then, patients were evaluated for the presence of HP infection and serum adiponectin level. After 8-hour fasting overnight, blood samples were taken at 4°C and, right after sampling, the serum was centrifuged at 2000 rpm for 15 minutes.Anti-HP IgG antibody was measured by ELISA kit, made by Padtan Elm Co, Iran. In case of serum titer above 30 AU/mL, it was considered positive. Serum adiponectin was used by AviBion Human Adiponectin ELISA-FINLAND kit based on ng/mL. Serum insulin was measured using ELISA kit, DiaMetra Co, Italy. Multiplying the fasting glucose value (mg/dL) by serum insulin value in each person and then dividing it by 405, HOMA-IR was calculated in this study.t-test, Chi-square test, and Fisher's exact test.Data analysis was performed by SPSS version 16 using descriptive statistics, mean, standard deviation, percentage, and frequency. The analysis was performed by From 211 patients enrolled in the study, 135 (64%) were female and 76 (36%) were male. The mean age and mean diabetes duration were 52.5 ± 8.8 and 7.4 ± 5.4 years, respectively.P = 0.427).In this study 139 patients were HP+ (65.87%) and the mean BMI in HP− and HP+ groups was 29.35 ± 4.9 and 28.79 ± 4.81, respectively, and it was not statistically significant (P = 0.778).In HP− group, 47 (65.3%) patients were female and 25 (34.7%) were male and in HP+ group, 88 (63.3%) patients were female and 51 (36.7%) patients were male; HP and gender had no statistically significant difference (P = 0.642). Also, blood pressure was measured in both groups and had no statistically significant difference (P = 0.825)  .P = 0.939). The mean chemical biomarkers like FBS in HP− and HP+ groups were 173.43 ± 61.31 mg/dL and 180.11 ±  64.28 mg/dL, respectively. Although it was higher in HP seropositive patients, it was not statistically significant (P = 0.468). Also, the mean HbA1c in HP− and HP+ groups was 8.08 ± 1.32 and 8.11 ± 1.67 mg/dL, respectively (P = 0.895).Family history of diabetes in HP− group was positive in 53 (73.6%) and in HP+ group, 103 (74.1%) had a positive familial history of diabetes (P = 0.140).The mean adiponectin level in HP− group was 4.54 ± 5.43, in HP+ group it was 5.64 ± 3.88 ng/mL, and it was not statistically different in both groups (P = 0.002) and insulin resistance degree was significantly higher in HP+ group . The mean AST was, respectively, 20.7 ± 10.07 and 22.9 ± 13.8 mg/dL in HP− and HP+ groups (P = 0.244). Also the mean ALT in HP− and HP+ groups was 21.8 ± 7.7 versus 23.8 ± 14.5 (P = 0.274). Creatinine in HP− group was 0.91 ± 0.19 and in HP+ group it was 0.95 ± 0.29 mg/dL (P = 0.589).Serum insulin in HP− and HP+ groups was 6.97 ± 5.64 versus 10.12 ± 7.72 (Helicobacter pylori is a human pathogen infecting the gastric mucosa which causes inflammatory process that increases in chronic gastritis, peptic ulcer, gastric lymphoma of mucosa-associated lymphoid tissue, and adenocarcinoma [arcinoma . Aydemirarcinoma .Adiponectin is a hormone protein secreted by adipose tissue and its functions in the body include regulation of blood glucose through gluconeogenesis reduction and increased glucose uptake by cells, catabolism of fatty acids, lipid catabolism, protecting vascular endothelial dysfunction, increased insulin sensitivity, weight loss, and control of energy metabolism .In most of the studies like Lihn et al.'s study, the low level of adiponectin is considered an important factor in development of metabolic syndrome and diabetes. They reported that the serum adiponectin level of patients with metabolic syndrome was significantly lower than patients without metabolic syndrome . Also, OAlso, Weyer et al. reported that hypoadiponectinemia has a close association with insulin resistance and hyperinsulinemia and inflammatory factors are involved in the regulation of adiponectin .Hung et al.'s study in 2008 evaluated the association of adiponectin levels with inflammatory markers, insulin resistance, and metabolic syndrome in 1094 patients 27–77 years old. They found that plasma adiponectin was a significant negative correlate of the levels of CRP and IL-6. Plasma adiponectin was also an inverse associate of HOMA-IR .In our study, we found that, although HP infection increases insulin resistance in diabetic patients, this organism does not make a significant change in adiponectin level and it may aggravate insulin resistance in diabetic patients from other mechanisms. Few studies have been conducted to assess HP infection and serum adiponectin. In Ando et al.'s study in 2012, the effect of HP eradication on serum adiponectin level was evaluated in 456 patients under endoscopy for HP diagnosis. Of these patients, 338 ones had HP and underwent treatment. The serum level of adiponectin had no statistically significant difference in HP positive and negative patients but HP eradication increased the adiponectin level .In another study by Hainer et al., low adiponectin conferred independent risk for impaired insulin sensitivity along with elevated WCC and HP titer .HP seropositive diabetic patients have higher insulin resistance degrees but the adiponectin serum level in these patients has no statistically significant difference with seronegative patients and it seems that increased insulin resistance has another reason in these patients.


Annona macroprophyllata  leaves was evaluated on both the activity of yeast α-glucosidase (an in vitro assay) and sucrose tolerance in Wistar rats. The results have shown that the aqueous extract from A. macroprophyllata inhibits the yeast α-glucosidase with an IC50 = 1.18 mg/mL, in a competitive manner with a Ki = 0.97 mg/mL, a similar value to that of acarbose (Ki = 0.79 mg/mL). The inhibitory activity of A. macroprophyllata was reinforced by its antihyperglycemic effect, at doses of 100, 300, and 500 mg/kg in rats. Chromatographic analysis identified the flavonoids rutin and isoquercitrin in the most polar fractions of A. macroprophyllata crude extract, suggesting that these flavonoids are part of the active constituents in the plant. Our results support the use of A. macroprophyllata in Mexican folk medicine to control postprandial glycemia in people with diabetes mellitus, involving active constituents of flavonoid nature.Annona genus contains plants used in folk medicine for the treatment of diabetes. In the present study, an aqueous extract prepared from  Diabetes mellitus (DM) is a chronic disease associated with abnormal and constant high blood glucose (hyperglycemia) that results from defects in insulin secretion, action, or both. In type 2 diabetes mellitus (DM2), postprandial hyperglycemia is important in the development of the disease; it is associated with micro- and macrovascular complications, and it has been proposed as an independent risk factor for cardiovascular disease , 2. The α-glucosidases hydrolyze starch and oligosaccharides [α-glucosidases, exoglycosidases, found in the luminal surface of enterocytes contain maltase/glucoamylase and sucrose/isomaltase activities [α-glucosidase activity [α-glucosidase inhibitors, such as acarbose, have been used in the clinic to control blood glucose increase, especially postprandial, in DM2 [α-glucosidases but have the disadvantage of undesirable side effects such as abdominal distention, flatulence, meteorism, and diarrhea [α-amylase, resulting in the abnormal bacterial fermentation of undigested carbohydrates in the colon. Therefore, more effective inhibitors for the enzymes should have strong inhibitory effect against α-glucosidase and mild inhibitory effect against α-amylase, which can be an effective therapy for managing postprandial hyperglycemia with minimal side effects [One important factor that arises in postprandial hyperglycemia is the fast uptake of glucose in the intestine, in which charides . The α-gtivities . It is bactivity . The α-g, in DM2 ; these cdiarrhea . Previou effects .Annona, Rollinia, Uvaria, and Asimina) produce edible fruits such as annona [Annona macroprophyllata Donn. Sm. is a species belonging to the Annona genus; it is a common tree in central México known as “ilama” ; whereas in the southeast (Tehuantepec region and Yucatán) it is called “papauce” or “anona blanca” [Annona squamosa, A. muricata, A. glabra, and A. cherimola [A. macroprophyllata, besides its described medicinal properties [Annonaceae is a family of plants consisting of 2300 to 2500 species included in more than 130 genera; in fact, it is the largest family of the order Magnoliales. Only four genera , certified the authenticity of the plant, and a voucher specimen (AN9702) was deposited in the herbarium “Herbario de Plantas Útiles Efraím Hernández X” at the UACH, in the State of México, México, for future reference. Aqueous extract was obtained by using 50 g of dried and powdered leaves, in a process of infusion in 500 mL of boiling water. The aqueous extract was separated from the residues by gravity filtration; samples were frozen in liquid nitrogen and then lyophilized during 12 h using a Heto FD3 Lab lyophilizer to yield 1.11 g (2.22%). μm particle size). The samples were injected through a 20 μL loop. The column was thermostatically controlled at 30°C using a 1 mL/min flow rate. The mobile phase consisted of 15 : 85 acetonitrile-trifluoroacetic acid solution at pH 2.5. The detection was monitored at 350 nm wavelength, and the system was run for 15 min. Calibration curves were done for the standards: kaempferol, hesperidin, naringenin, naringin, rutin, isoquercitrin, and quercetin.An HPLC analysis was performed using an Agilent Technologies Chromatograph. The separation was performed on an ODS Hypersil C18 column (15 mm × 4 mm i.d. and 5 α-Glucosidase inhibition was measured at pH 7.0 and 30°C using p-nitrophenyl-α-D-glucopyranoside (pNPG) as substrate and 0.75 IU/mL of yeast α-glucosidase in 0.1 M sodium phosphate buffer. Acarbose and aqueous extract were dissolved in phosphate buffer, and serial dilutions from 1.4 to 0.2 mg/mL were prepared. The increments in absorption at 405 nm, due to the hydrolysis of pNPG by the α-glucosidase, were determined on a microplate reader DTX 880 Multimode Detector from Beckman Coulter. Ten μL of acarbose or extract solution (in triplicate) were incubated during 5 min with 20 μL of enzyme stock. After incubation, 10 μL of substrate were added and further incubated for 35 min at 30°C. Finally, the reaction was stopped by adding 30 μL of 5 M Na2CO3. The concentration required to inhibit the enzyme activity by 50% (IC50) was calculated by regression analysis, using the following equation:v is the percentage of inhibition, A100 is the maximum inhibition, I is the inhibitor concentration, IC50 is the concentration required to inhibit the enzyme activity by 50%, and s is the cooperative degree.α-glucosidase was determined by the Lineweaver-Burk plots. All the results are expressed as the mean of at least three experiments ± SEM. Kinetic parameters such as Vm, Km, and Ki were evaluated by using the nonlinear regression method, based on the following inhibition equation:v is the initial velocity in either the Presence or absence of the inhibitor S and I are the concentrations of substrate and inhibitor, respectively; Vm is the maximum velocity, Km is the Michaelis-Menten constant, Ki is the competitive inhibition constant, and Ki′ is the uncompetitive inhibition constant. The kinetic data were analyzed using a computer program for nonlinear regressions (Origin 8.0).The mode of inhibition for ad libitum.Male Wistar normoglycemic rats, weighing 200–250 g, were obtained from our animal facilities at Facultad de Estudios Superiores Iztacala, U.N.A.M. Procedures involving animals and their care fulfilled the Mexican Official Norm for Animal Care and Handling (NOM-062-ZOO-1999) and were in compliance with international rules on care and use of laboratory animals. Furthermore, clearance for conducting the studies was obtained from the Ethics Committee for the Use of Animals in Pharmacological and Toxicological Testing of Facultad de Estudios Superiores Iztacala, U.N.A.M. For the pharmacological studies, groups of four animals were used. All doses are in mg/kg of body weight. The rats were housed in groups of four under standard laboratory conditions (12 h light/dark cycle at 22 ± 1°C) and maintained on a standard pellet diet and water Gi is the initial glycemia values and Gt is the glycemia value after treatments administration [All aqueous administrations were suspended in 0.9% saline solution and given by intragastrical route. Acarbose  was used as antihyperglycemic drug. Sucrose  was used as carbohydrate to carry out the sucrose tolerance tests. The control rats received only vehicle  in the same volume (0.5 mL of vehicle/100 g of body weight) by the same route. Blood samples were collected from the caudal vein by performing a small incision at the end of the tail. Blood glucose (mg/dL) was estimated using a commercial glucometer . The variation of glycemia as percentage for each group was calculated with respect to the initial (0 h) level, according to the following equation:stration .A. macroprophyllata was tested at the doses of 100, 300, and 500 mg/kg. Acarbose (5 mg/kg) was suspended in the same vehicle. Time 0 min was set before treatment with the extract; 30 min later a sucrose load (3 g/kg) was administered to the rats. Blood samples were obtained 15, 30, 60, 90, and 120 min after the carbohydrate load [Rats were fasted during 12 h before the experiment having free access to water. The crude extract of ate load .n = 4 rats for the OSTT and n = 3 assays for in vitro experiments. Differences were analyzed using Student's t-test (Sigma Stat version 3.0), and significant differences were considered at P < 0.05.Data represent the mean ± SEM of A. muricata, A. glabra, and A. cherimola as antidiabetic species [A. macroprophyllata is an interesting species to study, since it may have the same metabolic pathways that synthesize the compounds as the other members of the genus and thus may show their therapeutic properties. That is, we tested a chemotaxonomic criterion for the study.In México, it has been reported the use of  species , which sA. macroprophyllata leaves aqueous extract yielded rutin  and isoquercitrin , as well as some traces of astragalin/isoquercitrin (0.03 μg/mg of crude extract) (Elution of extract) . Annona macroprophyllata ethanolic extract has been investigated as antinociceptive, anti-inflammatory, and anticonvulsant agents [α-glucosidase activity both in vivo and in vitro [t agents –17; howet agents , 22–25. t agents . Rutin ht agents , and rutt agents –30. Rutiin vitro , and thein vitro .A. macroprophyllata inhibited the activity of yeast α-glucosidase, with a low IC50 (1.18 mg/mL), very similar to that of acarbose (0.27 mg/mL). This result agrees with that reported for rutin as inhibitor of α-glucosidase (IC50 = 0.196 mM), compared with acarbose (IC50 0.091 mM) [A. macroprophyllata on the α-glucosidase inhibition and antihyperglycemic effects.To our knowledge, this is the first report that the aqueous extract from .091 mM) ; therefoA. macroprophyllata, then a kinetic analysis of the inhibition of enzyme activity using different amounts of aqueous extract or acarbose was conducted. The results yielded a Ki of 0.97 mg/mL and Ki of 0.79 mg/mL to aqueous extract and acarbose, respectively. These values show that the extract inhibited the enzyme and could be used to control postprandial hyperglycemia. Lineweaver-Burk plots for α-glucosidase from yeast, in the presence of acarbose or A. macroprophyllata extract at different concentrations . The postprandial blood glucose peak was diminished from 15 min in all three doses ; however, the highest activity was 300 > 500 > 100 mg/kg throughout the time-curve postingestion of sucrose. The glycemic lowering effect of A. macroprophyllata at doses of 100, 300, and 500 mg/kg was not greater than that with acarbose, a therapeutic drug used as positive control (5 mg/kg); however, maximum effect obtained at a dose of 300 mg/kg was not statistically different with acarbose dose from 60 min on, results in vivo that agree to the reported for other species of the genus Annona as is the case of A. squamosa, A. muricata, and A. cherimola, which also show antioxidant, hypoglycemic, antihyperglycemic, and hypolipidemic properties [A. macroprophyllata involves an antihyperglycemic effect, mediated by the regulation of glucose uptake from the intestinal lumen, through the inhibition of carbohydrate digestion, putatively by inhibition of intestinal α-glucosidases complex, as observed in vitro for yeast α-glucosidase. In addition, the search for the active principle of the extract could generate a new α-glucosidase inhibitor, useful for the development of new antidiabetic or antiobesity agents.On the other hand, to confirm whether our results operties , 22–25. A. macroprophyllata aqueous extract has potent antihyperglycemic effect, and according to the in vitro studies it acts as a competitive inhibitor of the α-glucosidase. Last but not least, this study provides scientific support to use A. macroprophyllata leaves in Mexican traditional medicine for the treatment of DM2.In conclusion, our results indicate that


The corrosion of steel in concrete is mainly due to the chemical reaction between the chloride ions and iron ions. Indeed, this is a serious threaten for reinforced concrete structure, especially for the reinforced concrete structure in the sea. So it is urgent and important to protect concrete against chloride ions corrosion. In this work, we report multilayer concrete can cloak chloride ions. We formulated five kinds of concrete A, B, C, D and E, which are made of different proportion of cement, sand and glue, and fabricated six-layer (ABACAD) cylinder diffusion cloak and background media E. The simulation results show that the six-layer mass diffusion cloak can protect concrete against chloride ions penetration, while the experiment results show that the concentration gradients are parallel and equal outside the outer circle in the diffusion flux lines, the iso-concentration lines are parallel outside the outer circle, and the concentration gradients in the inner circle are smaller than those outside the outer circle. In 2006, Pendry designed perfect invisibility cloak234567891011121314151618192021222526283031323334353637t is time, C is mass diffusion concentration , D is diffusion coefficient (m2 · s−1), x, y, z, t), i.e., C = C . According to the mass diffusion theory, we can obtain the following simplified radial and tangential mass diffusion coefficient similar thermal diffusion coefficient33AD and BD, respectively, the thicknesses are Ad and Bd, respectively, we obtain the following effective parameters rD and θDrD is the radial diffusion coefficient, and θD is the tangential diffusion coefficient.The mass diffusion equation can be written as: c = Db = 1.0), the diffusion coefficient of the hollow cylinder may be an effective diffusion coefficient of isotropy and homogeneous composite material, the diffusion flux lines are parallel both in the background media and hollow cylinder, this phenomenon is called “mass diffusion transparency”. c = 4.0) is larger than that of the background media (Db = 1.0), the diffusion flux lines are attracted by the hollow cylinder, and the diffusion flux lines are distorted both inside and outside the hollow cylinder. c = 10000) is much larger than that of the background media (Db = 1.0), the diffusion flux lines are completely attracted by the hollow cylinder, no diffusion flux lines penetrate into the inner circle, this phenomenon is called “mass diffusion shielding”, just like magnetostatic (electrostatic) shielding is generated by large permeability (permittivity) material. c = 0.25) is smaller than that of the background media (Db = 1.0), the diffusion flux lines are repelled by the hollow cylinder, and the diffusion flux lines are also distorted both inside and outside the hollow cylinder. c = 0.0001) is much smaller than that of the background media (Db = 1.0), the diffusion flux lines are completely repelled by the hollow cylinder, no diffusion flux lines penetrate into the inner circle, just like magnetic field lines are completely repelled by an “superconductor”. According to the diffusion theory, we simulate mass diffusion. A = 4.000 × 10−8 cm2/s, DB = 0.278 × 10−8 cm2/s, DC = 0.547 × 10−8 cm2/s, DD = 0.741 × 10−8 cm2/s, respectively, the diffusion coefficient of the background media are DE = 1.000 × 10−8 cm2/s. 3 on the left plane, and the blue parts represent small concentration region, the concentration is set to be 0 on the right plane, the horizontal lines are diffusion flux lines (According to Eq. (2) and Eq. (3), we design six-layer (ABACAD) diffusion cloak and simulate concentration distributions. The inner and outer radii of the six-layer (ABACAD) are , ,  , , , respectively, and the diffusion coefficients of the four kinds of materials are Dux lines . The incA = 4.000 × 10−8 cm2/s, DB = 0.253 × 10−8 cm2/s, DC = 0.478 × 10−8 cm2/s, DD = 0.650 × 10−8 cm2/s, DE = 0.766 × 10−8 cm2/s, DF = 0.855 × 10−8 cm2/s, respectively, the diffusion coefficients of the background media are DG = 1.000 × 10−8 cm2/s. The diffusion coefficient of the background media are DE = 1.000 × 10−8 cm2/s. Simulation results show that the more the layers, the better performance the mass diffusion cloak, and the small diffusion coefficient materials exclude diffusion flux lines and the large diffusion coefficient materials attract diffusion flux lines, just like small permittivity (permeability) material excludes electric (magnetic) field line and large permittivity (permeability) material attracts electric (magnetic) field line. It should be pointed out that the mass diffusion cloak mainly acts as a flux-guiding shell to suppress the influence of diffusion reflection.b = 1.000 × 10−8 cm2/s and Dc = 2.500 × 10−8 cm2/s, respectively. Outside the outer circle, the iso-concentration lines are not parallel and the horizontal diffusion flux arrows are also not parallel. So this hollow cylinder is a non-cloak.Utilizing the transient diffusion equation of COMSOL Multiphysic software, we simulate the diffusion cloak in different time. The parameters are the same as that in A = 4.00 × 10−8 cm2/s, DB = 0.28 × 10−8 cm2/s, DC = 0.55 × 10−8 cm2/s, DD = 0.74 × 10−8 cm2/s, DE = 1.00 × 10−8 cm2/s, respectively. According to the measurement results, we find that the more the glue, the smaller the diffusion coefficient, i.e., the glue can decrease diffusion coefficient. It is also proved that other material, such as fly ash, can also decrease diffusion coefficient. According to the simplified parameter Eq. (2) and effective parameter Eq. (3), we fabricate the six-layer (ABACAD) cylinder diffusion cloak. The inner and outer radii of the six-layer cylinder cloak are , , , , , , respectively, the height of the cylinder cloak is h = 6 cm . After 100 minutes diffusion, pasting simulation results . In the inner circle, the test points are located at  and , respectively, outside the outer circle, the test points are located at  and , respectively. The concentration gradients in the inner circle are always smaller than those outside the outer circle in the cloak even after 550 minutes . The conGuenneauFor an ideal mass diffusion cloak, the diffusion matters do not penetrate into the inner circle, thus a perfect diffusion protection is offered. The multilayer mass diffusion cloak is a simplified approximate cloak. The chloride ions can penetrate into the cloak region in long time diffusion. In a steady state, the cloak region eventually achieves a constant concentration. Therefore, the mass diffusion cloak can only provide a temporal diffusion protection rather than a permanent protection. Another case is that the diffusion matter chloride ions pollution would change the effective parameters of the cloaking materials; finally the chloride ions go into the inner cylinder after long time diffusion. In our experiment, we chose 0.5% NaCl solution. The chloride ions concentration in point P  of Chloride ions pollution will corrode rebars. Controlling chloride ions diffusion in concrete has a great significance in the field of building material. Concrete is composed of cement, aggregate and paste interfacial zone404243We numerically simulate the steady and transient mass diffusion cloak by COMSOL Multiphysic software. The parameters fulfill Eq. (2) and Eq. (3).The samples are fabricated by six-layer concretes (ABACAD) and background media E, the concretes are made of cement, sand and glue mixture. The proportion of cement, sand and glue are A (1:3:0.02), B (1:3:0.30), C (1:3:0.20), D (1:3:0.10), E (1:3:0.08), respectively.NJCLChloride Ions Concentration Fast Measurement Equipment.According to the simulation results of time-dependent mass diffusion cloak, we dug up a lot of small cylinders in the iso-concentration lines and concentration gradient lines, and measured the concentration of chloride ions by L.Z. wrote the paper and numerical stimulated. R.S. set up the experiment and revised the manuscript.


Dental amalgam is a source of elemental and inorganic mercury. The safety of dental amalgam in individuals remains a controversial issue. Urinary mercury concentrations are used to assess chronic exposure to elemental mercury. At present, there are no indications of mercury-associated adverse effects at levels below 5 μg Hg/g creatinine (Cr) or 7 μg Hg/L (urine). The purpose of the present study is to determine the overall urinary mercury level in the Canadian general population in relation to the number of dental amalgam surfaces.Data come from the 2007/09 Canadian Health Measures Survey, which measured urinary mercury concentrations in a nationally representative sample of 5,418 Canadians aged 6–79 years. Urinary mercury concentrations were stratified by sex, age, and number of dental amalgam surfaces.th percentile was 2.95 μg Hg/L, the 99th percentile was 7.34E μg Hg/L, and the 99.9th percentile was 17.45 μg Hg/L. Expressed as μg Hg/g Cr, the overall 95th percentile was 2.57 μg Hg/g Cr, the 99th percentile was 5.65 μg Hg/g Cr, and the 99.9th percentiles was 12.14 μg Hg/g Cr. Overall, 98.2% of participants had urinary mercury levels below 7 μg Hg/L and 97.7% had urinary mercury levels below 5 μg Hg/g Cr. All data are estimates for the Canadian population. The estimates followed by the letter “E” should be interpreted with caution due to high sampling variability (coefficient of variation 16.6%-33.3%).The overall mean urinary mercury concentration varied between 0.12 μg Hg/L and 0.31 μg Hg/L or 0.13 μg Hg/g Cr and 0.40 μg Hg/g Cr. In general, females showed slightly higher mean urinary mercury levels than men. The overall 95The mean urinary mercury concentrations in the general Canadian population are significantly lower than the values considered to pose any risks for health. Mercury occurs in nature as elemental, inorganic, and organic mercury  . In termIn vitro studies have shown that the rate and amount of mercury released depends on the age and the condition of the amalgam restoration; the formation of a passive tarnish layer on the surface of the restoration is believed to be associated with a decrease in mercury vapour release .The variables for this analysis included: sex , age, and number of amalgam surface. The analysis was conducted for females, males and the two sexes combined, and for the following age intervals: 6–11 years, 12–19 years, 20–39 years, 40–59 years, 60–79 years, 6–79 years, and 16–49 years .Under the Statistics Act, Statistics Canada is required to ensure respondent confidentiality. Therefore, estimates based on a small number of respondents are suppressed. Estimates of the geometric mean require at least 10 respondents. Estimates at the 95th percentile require at least 200 respondents .The results are presented both as μg Hg/g Creatinine and μg Hg/L in order to facilitate comparison with other studies. Geometric means were determined because they are less influenced by extreme values, providing a better estimate of central tendency .Statistical analyses were based on weighted data, and were descriptive. To account for survey design effects, standard errors, coefficients of variation and 95% confidence intervals were estimated using the bootstrap technique. Specifically, a survey weight is given to each person included in the final sample. This weight corresponds to the number of persons represented by the respondent for the entire population. When an estimate is produced, the survey weight must be used to ensure the estimate represents the population. If the survey weight is not used, the estimate represents only the sample .In order to determine the quality of an estimate using the coefficient of variation (CV) or to calculate confidence intervals, the standard error of the estimate must be calculated. Since the CHMS uses a multi-stage survey design, there is no simple formula that can be used to calculate sampling variance. Instead, the bootstrap re-sampling method is used to take into account the sample design information and to easily obtain variance estimates. This method selects in each stratum, a simple random sample of (n-1) of the n first stage sampling units selected with replacement to form a replicate. In each replicate, the survey weight for each record in the (n-1) selected first stage sampling units is recalculated. These weights are then post-stratified according to demographic information in the same way as the survey weights in order to obtain the final bootstrap weights. This process is repeated 500 times for the CHMS .All data were directly entered into a computer at the time of collection. For the oral health clinical examination, a detailed quality checking protocol was built into the data-entry program. With the extensive input of the Office of the Chief Dental Officer staff, Statistics Canada programmed entry values such that many areas of logical inconsistency were “greyed-out” and erroneous entries could not be made .All analyses were conducted in-house by Statistics Canada. An experienced methodologist also verified the coding to produce all estimates in an effort to ensure reliability of the estimates. Furthermore, all numbers were verified prior to publication. The tables were formatted to display the CHMS findings in such a manner that both took advantage of the rich information and also would allow readers to compare the findings with those of the other publications .The estimates are representative for the Canadian population (based on weighted data). All estimates accompanied by the letter “E” must be interpreted with caution due to high sampling variability; coefficient of variation 16.6% - 33.3%.In the amalgam free group, selected percentiles  and mumc estimates are provided for each age group . In the Overall group, the number and percentage of people with urinary mercury concentrations above 7 μg Hg/L and 5 μg Hg/g Cr are presented in addition to the above   had a mumc of 4.77E μg Hg/g Cr, 95% CI (2.43 - 9.35). Only 2 children  had 26–30 amalgam surfaces. When measured as μg Hg/g Cr, 2 children 6–11 years of age in the CHMS sample were identified as having urinary mercury levels above HBM I values. Estimates of the Canadian population represented and the corresponding percentages could not be provided due to insufficient data for analysis. Estimated as μg Hg/g Cr, 15 children in the CHMS sample, representing approximately 57,035 Canadians, or less than 2.64% of the child population (6–11 years of age) had urinary mercury levels above HBM I values  for which the mumc could be determined , had a mumc of 3.79E μg Hg/L, 95% CI (1.89 - 7.62) or 2.39E μg Hg/g Cr, 95% CI (1.42 - 4.01). There were7 adolescents  with 16–20 amalgam surfaces. Three adolescents in the CHMS sample had urinary mercury levels above HBM I values. Estimates of the Canadian population represented and the corresponding percentages could not be provided due to insufficient data for analysis  had a mumc of 0.84 μg Hg/L, 95% CI (0.81 - 0.87) or 2.03 μg Hg/g Cr, 95% CI (2.030 - 2.034). There were 2 individuals  with 46–65 amalgam surfaces. When measured as μg Hg/L, 13 people 20–39 years of age in the CHMS sample, representing approximately 293,798 Canadians, or less than 3.26% of the population (20–39 years of age) were identified as having urinary mercury levels above HBM I values. Estimated as μg Hg/g Cr, 10 people in the CHMS sample, representing approximately 159,516 Canadians, or less than 1.77% of the population (20–39 years of age) had urinary mercury levels above HBM I values  had a mumc of 3.34E μg Hg/g Cr, 95% CI 1.72 - 6.50) . There were 4 individuals  with 46–65 amalgam surfaces. When measured as μg Hg/L, 19 people 40–59 years of age in the CHMS sample, representing approximately 328,642 Canadians, or less than 3.37% of the population (40–59 years of age) were identified as having urinary mercury levels above HBM I values. Estimated as μg Hg/g Cr, 28 people in the CHMS sample, representing approximately 446,642 Canadians, or less than 4.58% of the population (40–59 years of age) had urinary mercury levels above HBM I values  also had the largest mumc, measured at 2.78E μg Hg/L, 95% CI 1.77 - 4.37) or 3.00E μg Hg/g Cr, 95% CI (1.82 - 4.95). There were 7 individuals  with 46–65 amalgam surfaces. When measured as μg Hg/L, 12 people 60–79 years of age in the CHMS sample, representing approximately 115,519 Canadians, or less than 2.35% of the population (60–79 years of age) were identified as having urinary mercury levels above HBM I values. Estimated as μg Hg/g Cr, 15 people in the CHMS sample, representing approximately 141,572 Canadians or less than 2.88% of the population (60–79 years of age) had urinary mercury levels above HBM I values  was 3.86E μg Hg/L, 95% CI (1.98 - 7.51) or 2.88E μg Hg/g Cr, 95% CI (1.65 - 5.03). There were 27 women with 31–45 amalgam surfaces. When measured as μg Hg/L, 20 women 16–49 years of age in the CHMS sample, representing less than 4.37% of the Canadian female population (16–49 years of age), were identified as having urinary mercury levels above HBM I values. Estimated as μg Hg/g Cr, 19 women in the CHMS sample, representing less than 4.74% of the Canadian female population (16–49 years of age) had urinary mercury levels above HBM I values  at least 96.63% - 98.16% had urinary mercury levels below 7 μg Hg/L, and at least 95.42% - 98.23% had urinary mercury levels below 5 μg Hg/g Cr . At most, 3.37% - 1.84% and 4.58% - 1.77% respectively had estimates that should be verified by further measurements (between HBM-I and HBM-II) [In Canada, the mumc in people with or without dental amalgam restorations are well below the levels associated with any health risks. In general, the mumcs tend to increase with the number of amalgam surfaces, and appear to be influenced by age and sex. On average, females showed slightly higher levels of urinary mercury than males. These observations are consistent with the findings of other researchers . HoweverFor the same exposure level, organic mercury from food results in an absorption approximately six-times greater than the absorption of inorganic mercury from dental amalgam restorations . The CHMFish consumption can elevate mumcs; once ingested, some of the methylmercury from fish is de-methylated to inorganic mercury, and excreted in urine ,22-25. TElemental, inorganic, and organic mercury compounds have specific half-lives and metabolic pathways. The high sampling variability reflects mercury ubiquity and its diverse metabolic pathways, which influence the uptake, distribution, accumulation patterns and excretion rates. The same baseline mercury exposure was assumed for all survey participants ; it is unknown if it was a correct assumption. For instance, one might argue that individuals with amalgam restorations have a different level of exposure from other sources than people without such restorations.The results of this research indicate that the overall mumc in the Canadian general population is two times lower than the overall mumc reported by the 2003–2004 NHANES . Furtheret al. found that people with bruxism and/or who were heavy gum chewers had urinary mercury levels ranging from 25 μg Hg/g Cr to 54 μg Hg/g Cr [th percentile estimate provided was 12.87E μg Hg/g Cr, 95% CI (6.7 - 19.03). Although high, this 99.9th percentile value is not associated with adverse health effects. Yet it indicates that some people require further investigation [In a number of case studies, Barregard  Hg/g Cr . Such ur Hg/g Cr . The restigation . Additioet al.[In people without dental amalgam, the difference between the estimates provided by Barregard et al. and the et al..Although most studies on the subject report mumcs, it is important to look beyond the average . SelecteA recent study explored various scenarios of mercury exposure and risks from dental amalgam in the US population. According to the worst case scenario more than 60 million Americans would exceed the Hg reference exposure level established by the US Environmental Protection Agency . HoweverWhile a worldwide ban of dental amalgam will not impact on total worldwide mercury pollution , a ban oDisease rates are higher in the poorest and the most vulnerable populations; such cost increases would arguably concentrate in these already disadvantaged groups. Using population projections to obtain national estimates of dental amalgam use, and the dental component of the Consumer Price Index to estimate the annual rate of change in fees, it has been estimated that the first-year of banning dental amalgam in the entire population in the U.S. would lead to an $8.2 billion increase in direct dental expenditures alone .Health Canada points out that current evidence does not indicate that dental amalgam is causing illness in the general population . Also, aImportantly, dental amalgams are usually replaced with composite resin restorations, which are a source of BPA . The EurFor the vast majority of Canadians, the mean urinary mercury concentrations are below a level of concern regardless of the number of amalgam surfaces they have. It is arguable that from a population health standpoint, dental amalgam remains a safe restorative material.Written informed consent was obtained from the patient’s guardian/parent/next of kin for the publication of this report and any accompanying images.CHMS: Canadian health measure survey; CTQ: Centre de toxicologie du Québec; Cr: Creatinine; Hg: Mercury; INSPQ: L'Institut national de santé publique du Québec; NHANES: National health and nutrition examination survey; Mumc: Mean urinary mercury concentration; Mg Hg/L: Micrograms mercury per liter (in urine); Mg Hg/g: Cr micrograms mercury per gram creatinine (in urine).The authors declare that they have no competing interests.AN participated in the design of the study, results interpretation and helped to draft the manuscript. HA conceived of the study, and participated in its design and coordination and helped to draft the manuscript. CQ participated in the design of the study and helped to draft the manuscript. All authors read and approved the final manuscript.AN, DDS, MSc, FRCD(C), HA, DDS, MPH, Membership Services Coordinator, Alberta Dental Association.CQ, DMD, MSc, PhD, FRCD(C) Assistant Professor and Director, Specialty Training Program  Dept. of Biological and Diagnostic Sciences/Community Dentistry, University of Toronto.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6831/13/44/prepub