Background. Microarray technology shows great potential but previous studies were limited by small number of samples in the colorectal cancer (CRC) research. The aims of this study are to investigate gene expression profile of CRCs by pooling cDNA microarrays using PAM, ANN, and decision trees (CART and C5.0). Methods. Pooled 16 datasets contained 88 normal mucosal tissues and 1186 CRCs. PAM was performed to identify significant expressed genes in CRCs and models of PAM, ANN, CART, and C5.0 were constructed for screening candidate genes via ranking gene order of significances. Results. The first screening identified 55 genes. The test accuracy of each model was over 0.97 averagely. Less than eight genes achieve excellent classification accuracy. Combining the results of four models, we found the top eight differential genes in CRCs; suppressor genes, CA7, SPIB, GUCA2B, AQP8, IL6R and CWH43; oncogenes, SPP1 and TCN1. Genes of higher significances showed lower variation in rank ordering by different methods. Conclusion. We adopted a two-tier genetic screen, which not only reduced the number of candidate genes but also yielded good accuracy (nearly 100%). This method can be applied to future studies. Among the top eight genes, CA7, TCN1, and CWH43 have not been reported to be related to CRC. Colorectal cancer is one of the leading cancers in the world and considered to be among the most frequent causes of death, along with lung, prostate, and breast cancer . MicroarThe study pooled the dataset of microarrays from different research teams in the Gene Expression Omnibus (GEO) database in order to increase sample size, sample heterogeneity, and statistical power in the hope of addressing the issue of insufficient sample size presented in previous studies. Four methods, Prediction Analysis of Microarray (PAM), Artificial Neural Network (ANN), Classification and Regression Trees (CART), and C5.0, were employed to analyze the variations in gene expression between colorectal tumors and normal mucosa tissues in order to screen for significant genes.The microarray gene expression data are from searches using “colon cancer” and “human [organism]” and “expression profiling by array [dataset type]” as the keywords in the GEO database of the National Center for Biotechnology Information (NCBI). The eligible criteria were as follows: (1) the examined samples were frozen tissue sections of normal human colorectal mucosa, primary colorectal cancer, or hepatic metastases from colorectal cancer; (2) the microarray platform used was limited to single-color, whole genome gene chips from Affymetrix; and (3) the data were presented as gene expression level. The exclusion criteria were as follows: (1) data from cultured cell lines or other in vitro assays; (2) datasets without the original gene expression level data files; and (3) those with redundant subdatasets. A total of 190 GEO series (GSE) datasets were finally 174 excluded, leaving 16 independent datasets for analysis, which are as follows: GSE4045, GSE4107, GSE4183, GSE5851, GSE8671, GSE9348, GSE1096, GSE12630, GSE12945, GSE13067, GSE13294, GSE13471, GSE15960, GSE17538, GSE18105, and GSE14333. The total number of examined tissues was 1,274, including 88 normal mucosal tissues and 1,186 colorectal tumor tissues . The data set is unbalanced among groups of adenoma, adenocarcinoma, carcinoma, metastasis, and normal tissues having 53, 521, 533, 79, and 88 samples, whereas, in training set, candidate genes modeling two remaining groups between cases and controls must have accuracies more than 88/(88 + 53) = 0.6241, 521/(88 + 521)= 0.8555, 533/(88 + 533) = 0.8583, and 88/(88 + 79) = 0.5269, respectively.All microarray data of examined tissues were produced from primary surgical cases without chemotherapy. For adjusting metastatic spread, all the cases were divided into five groups based on pathological tissue types: normal mucosal tissues (nm), adenoma (ad), adenocarcinoma (ac), primary tumor , and liver metastases (mt). More clinical factors of stage, grade, microsatellite instability (MSI), microsatellite stability (MSS), sites, race, gender, and age were adjusted for gene ordering and secondary genetic screening in the multinomial and multivariate logistic regression. The research process of flow chart diagram for the pooled datasets with analysis included in the study is shown in Figures In For instance, the HG-U133A gene chip used in this study is comprised of 22,283 probes that cover 14,713 genes. Each gene is covered by 1–14 probes. Of the 14,713 genes, 5,107 (38%) are covered by more than two probe combinations. For genes covered by multiple probe combinations, this study adopted the median method. For example, when the expression level of the HFE gene was reflected by the levels of 13 probes, the level of the seventh (the median number) probe was chosen to represent the expression of the HFE gene.Affymetrix chips were HG-U133A, HG-U133A-2, and HG-U133-Plus-2. Among these 3 types of chips, the corresponding numbers of genes were 14,713, 14,704, and 33,727, after the conversion that each single gene can map several probes, respectively. In the present study, the quartile normalization of all gene expression values was performed before combining 16 GSEs. The probes were merged to obtain the expression levels of 14,698 genes. The normalization can eliminate the systematic variations among studies .The candidate genes of colorectal tumors and normal mucosal tissues were selected from the primary screening of the 14,698 genes using PAM, followed by the construction of a classification model, determination of gene order of importance, and secondary gene screening using ANN, CART, C5.0, and PAM. decision trees were constructed using CART and C5.0 for intragroup comparison of gene order of importance.The ad, ac, cn, and mt groups were subjected to pairwise comparison of gene expression with the nm group to identify genes that were differentially expressed at different cancer stages. The bootstrapping rounds were used to avoid the poor extrapolation of the selected candidate genes that the proposed analysis is unbiased and would not lead to over-fitting. Three-fourths of the samples in each pair of comparison groups were randomly selected as the model-training group for PAM, and the remaining fourth served as the model-testing group for an independent set of samples that is left for final validation. Also, it will be interesting to see the results of the current method under the null hypothesis of nor predictive signal, which can be achieved by label permutation. The threshold criteria for gene selection by PAM were that the candidate genes were with the minimum classification errors, which number was lower than 100.http://dx.doi.org/10.1155/2014/634123). The introduction of the four analysis methods is as follows. The Clementine 10.1 was performed for ANN, C5.0, and CART and the R was used for PAM.Using the significant genes from the primary screening, four classification models were constructed for colorectal tumor tissues  and normal mucosal tissues (nm) for the secondary screening of significant genes and determination of the gene order of importance. Completely separating training and validation data sets is a crucial step in classifier development . It is very important that none of the samples in the validation dataset has been used in any part of the training, for example, for feature selection. In PAM uses nearest shrunken centroids as its algorithm. It can eliminate noise signal , controlANN needs to go through repeated training and learning to construct good classification models. It has the following advantages: (1) it is suitable for the analysis of highly dimensional and uncommon data; (2) it can accept missing values and process noise of the information. Its limitations are as follows: (1) it is necessary to avoid over-training the model and (2) it has a nontransparent solution process known as a “black box” (3). ANN were trained by the setting items of method with quick back propagation, prevent overtraining with 80% samples, set random seed-seed with 720925, stop on with default, optimize with speed. The Quick back propagation with producing smaller hidden layer and random seed 720925 with randomization for avoiding over-fitting were default in ANN on Clementine 10.1.Decision trees are models with tree-like structures. Trimming the branches can solve the problem of over-training. In general, larger decision trees are more expressive and may have more predictive power, but the smaller a decision tree is, the stronger its simplicity is . The conThe expression values of the 55 genes were analyzed in the models constructed with ANN, CART, C5.0, and PAM, and the contribution level of each gene to the classification of colorectal tumors and normal mucosal tissues was obtained, which was designated as gene importance.CART, C5.0, and PAM used the number of times each gene was selected as a predictor in 1,000 bootstrapping rounds. A higher number of times indicated a higher importance for the gene. ANN listed the relative importance values of the input gene variables in the classification model, with larger values representing higher contribution levels. In addition to the ordering methods above, PAM and C5.0 also used the centroids values, as well as the location of a gene, as a node, in the decision tree, respectively, to calculate gene importance. The higher absolute values of centroids and a node, which are closer to the root of a decision tree , represent the higher importance for the gene. All the index levels of gene importance in each model were ranked and normalized into percentile ranked score (RS%). For precise settings, see the Supplementary Table  2.ANN: the ANN model listed the relative importance values (RI) of the input gene variables in the classification of colorectal tumors and normal mucosal tissues, with larger values representing higher contribution levels. The relative importance values were ranked with 1 point for the lowest value, 2 points for the next lowest value, and so on. Identical ranked scores (RS) were given to the genes with the same RI value. Finally, each gene's RS was divided by the highest RS in order to obtain the percentile ranked score (RS%). CART: the gene importance ordering method in the CART model was used to calculate the number of times each gene was selected as a node in the decision tree in the analysis with 1,000 repeated samplings. A higher number of times selected indicated a higher importance for the gene. The genes were ranked based on the number of times they were selected as a significant gene, with 1 point for the lowest number of times, 2 points for the next lowest number of times, and so on. The same ranked score (RS) was given to the genes with the same Sig value. Finally, the RS of each gene was divided by the highest RS to obtain the RS%. PAM: one of the PAM gene importance calculation methods is similar to CART. However, in addition to using Sig to calculate importance, PAM used the centroids values (Cen) to calculate gene importance. The detailed calculation method using Cen was as follows. The absolute values were obtained for the averages of the centroids from the PAM analysis results after taking 1,000 repeated samples for the four group pairs . Next, the sum of the absolute values for each gene was ranked, with 1 point for the lowest value, 2 points for the next lowest value, and so on. The same RS was given to identical centroids values. Finally, the RS of each gene was divided by the highest RS to obtain the RS%. C5.0: the calculation method is based on the number of times each gene is selected, similar to CART, as well as node location. The latter method was that when a gene, as a node, is closer to the root of a decision tree , it has a higher RS, while it has a lower RS, when it is closer to the tip of the branches, and so on. The RS% calculation method is the same as that of CART. The ranked scores were converted to percentiles in order to make it possible to compare the order of gene importance obtained from these four analysis methods.)The use of pooled GEO studies was secondary because only microarray data was available. Testing 55 genes in any experimental model would be beneficial for colon cancer biology. Therefore, the present study analyzed the functional pathways that are related to the tumor genesis of colorectal cancer using GSEA software version 2.07. The GSEA MSigDB provides a collection of annotated gene sets based on different sources of information, for example, gene ontology, pathways, or motifs . Input vThe age distribution indicated a higher proportion of colorectal cancer in people over 60 years old. No difference was found in the location of collected tissues or genders between the two groups .The PAM screens the candidate genes with highly statistical significances by bootstrapping and cross-validation and then classification algorithms implement models with selected genes that can discriminate between two groups among cancers and controls. The results demonstrate how consistent different methods of data mining perform and select candidate genes among ANN, decision trees, and PAM according to correlation coefficients via ranking statistical significances of 55 genes.Significant genes were found in the ad/nm, ac/nm, cn/nm, and mt/nm combination groups, respectively, and a total of 55 significant genes were identified . The repThese 55 genes are mainly localized and functional at the cell surface and at tight junction. Their molecular functions are related to transporter activity, binding, catalytic activity, enzyme regulator activity, and gelatinase activity. The biological processes that these genes are involved in mainly include biological adhesion, signaling, transporter, metabolic process, insulin secretion, and biological regulation (see the Supplementary Tables  5–7).The mean values for the test accuracy reached 0.97 or above in the classification of normal and colorectal cancer cases. ANN and C5.0 exhibited the best performances, and ANN showed the best model stability . CART reP < 0.001) (The results in < 0.001) . PAMΔ anP < 0.001). ANN showed a significant correlation with CARTΔ and C5.0Δ but had relatively low correlation coefficients in the range of 0.42–0.48 . All genes except TCN1 remained significant after individual controlling for gender, race, tissue location or age, and CA7, SPIB, and CWH43 were still significant when the four confounders were controlled for simultaneously. Except for SPP1, which did not show a correlation with microsatellite stability, the other genes all exhibited significant correlation after logistic regression of individual variables, with OR values ranging from 0.49 to 1.12. Of the eight genes, only SPP1 was significantly associated with cancer stage. The higher the expression level of SPP, the more advanced the cancer stage was (see the Supplementary Tables  9 and 10). The multivariate logistic regression was performed for acquiring odds ratios of 8 genes expression of each of the eight genes adjusted by demographics in the four different groups. These odds ratios allow comparisons of patterns of expression changes for each of the genes; for example, whether gene expression increases (odds ratio > 1) for groups with more severe lesions or whether there is only a difference between normal and disease tissue.In nd CWH43 . The RS% DEFA6, TCN1, and KLK11 are upregulated and NR3C1 and THRB are downregulated in adenoma, which can be earlier screening markers. The genes of ABCG2, CA7, HP, KIAA1199, and CLDN1 are upregulated and CA4, CHP2, CHST5, CLCA1, CLCA4, CLDN8, FAM55D, H3F3A, MUC2, and NR3C2 are downregulated in carcinoma and metastases, which can be prognosis monitoring markers. The markers are regulated consistently with same direction comparing the present study, Cardoso et al. 2007 and Chan et al. 2008 [The Supplementary Table  10 showed that AQP8, SPIB, SPP1, and TCN1 are GO biological process annotated as transporters; CA7 and GUCA2B are annotated as being involved in biological regulation and signaling, respectively; IL6R is annotated as being part of the immune system process, while no annotation was available for CWH43. As for GO molecular function annotation, CA7, IL6R, and SPP1 are annotated as having gelatinase activity; AQP8, SPIB, and TCN1 are annotated as being capable of binding; GUCA2B is annotated as a regulator of enzyme activity, and CWH43 is not annotated.Of these eight genes,The GO functional results were similar to the results from the GSEA analysis with the 55 genes, suggesting these 55 genes are indeed located in the significant functional pathways that affect the tumor genesis of colorectal cancer.The primary GO molecular function annotation is binding . Although peptide binding was downregulated in colorectal tumor tissues, the other types of binding were all upregulated. The activities of genes involved in channel regulation signal transduction and transportation were suppressed in colorectal cancer. Of the GO biological processes, cell cycle, DNA replication, DNA metabolic process, and the glutamate signaling pathway were upregulated in colorectal cancer, while response to stimulus (feeding behavior) and lipid metabolism was downregulated (see the Supplementary Table  11).The present study aimed at finding possible marker gene sets for colorectal cancer by using a two-step complex bioinformatical analysis. The in silico gene expression analysis results in a low numbered gene set that could be a potent classificatory set for CRC. In a biological point of view, the methods and the results are valid; however, the verification of the given gene set on an independent sample set would be necessary, even at protein level. Biostatistically, it would be interesting to know whether the selected, highly discriminative genes are also present in the excluded GSE datasets.Four models performed very well at classifying colorectal tumors and normal mucosal tissues with the average test accuracy rates above 0.97. ANN showed the best classification performance, with an average test accuracy of 0.99 ± 0.01. CART required the fewest genes, requiring an average of 1.7 genes to reach an average test accuracy of 0.97. C5.0 had the best accuracy with the fewest genes among the four methods.In general, except for C5.0 and CART, a poor consistency in gene importance rank ordering was observed among the different methods. Lee et al.  observed ABCG2, AQP8, CA1, CA4, CA7, CD177, CDH3, CLCA4, CLDN8, EDN3, FCGBP, GCG, GUCA2A, GUCA2B, KIAA1199, MMP7, MS4A12, MT1M, MUC2, PYY, SLC26A3, SLC4A4, SPIB, SST, and ZG16.Among 55 selected genes, 13 overlapped with significant genes mentioned in two review articles on gene expression analysis in colorectal cancer , 7. The CA7, SPIB, GUCA2B, AQP8, IL6R, SPP1, TCN1, and CWH4) and is related to tumor progress and metastasis. Its expression was elevated in advanced or metastatic tumor tissues [Gelatinase activity was one major GO annotation of these eight genes  and guanylin (GUCA2A) are very close in structure and biological functions. They are responsible for signal transduction that regulates the transport and secretion of liquids and electrolytes in the gastrointestinal tract during digestion , 26. The AQP8 expression was only found in the columnar surface of epithelial cells; little or no expression of AQP8 was observed in the colonic crypts or tumor cells [Aquaporin are responsible for water absorption and excretion in the gastrointestinal tract . They plor cells , which ior cells .IL6R is primarily involved in the immune response, inflammation, and hematopoiesis [opoiesis  and was opoiesis .Osteopontin protein can be used as a marker of tumor progression for breast cancer , lung caTCN1 is a regulator of the Wnt/beta-catenin pathway and enhances the expression of the target genes of beta-catenin, leading to cancer progression and outcome deterioration. In gastric cancer studies, TCN1 was significantly correlated with cancer stage, poor cell differentiation, lymph node metastasis, and a poor prognosis [rognosis .CWH43 has been reported to contribute to the cell wall integrity of Saccharomyces cerevisiae [ CWH43 was downregulated in colorectal tumor tissues (fold change = −4.59).revisiae . However CA7, TCN1, and CWH43 are novel genes, but have not been previously reported as related to colorectal cancer. Further researches will help us better understand their roles in colorectal cancer.This study adopted a two-tier genetic screen, which not only reduced the number of candidate significant genes but also resulted in an impressively nearly 100% test accuracy. This analysis method can be applied to future pooled microarray studies. The more important genes exhibit a more consistent ranking order among the different methods used.Supplementary materials contain the details about settings of models, parameters of selected genes, of optimizing selected number of genes and gene ontological pathways.


Based on Pearson coefficients between particle number concentrations and meteorological parameters, precipitation and temperature both had significantly negative relationships with particle number concentrations, whereas atmospheric pressure was positively correlated with the particle number concentrations. The diurnal variation of number concentration in nucleation mode particles correlated closely with photochemical processes in all four seasons. At the same time, distinct growth of particles from nucleation mode to Aitken mode was only found in spring, summer, and autumn. The two peaks of Aitken mode and accumulation mode particles in morning and evening corresponded obviously to traffic exhaust emissions peaks. A phenomenon of “repeated, short-lived” nucleation events have been created to explain the durability of high particle concentrations, which was instigated by exogenous pollutants, during winter in a case analysis of Wuhan. Measurements of hourly trace gases and segmental meteorological factors were applied as proxies for complex chemical reactions and dense industrial activities. The results of this study offer reasonable estimations of particle impacts and provide references for emissions control strategies in industrial cities of developing countries.Atmospheric fine particles (diameter < 1 μm) attract a growing global health concern and have increased in urban areas that have a strong link to nucleation, traffic emissions, and industrial emissions. To reveal the characteristics of fine particles in an industrial city of a developing country, two-year measurements of particle number size distribution (15.1 nm–661 nm), meteorological parameters, and trace gases were made in the city of Wuhan located in central China from June 2012 to May 2014. The annual average particle number concentrations in the nucleation mode (15.1 nm–30 nm), Aitken mode (30 nm–100 nm), and accumulation mode (100 nm–661 nm) reached 4923 cm Meglected ,16,17,18eglected , becauseeglected . Thus, ieglected .Extensive studies on PSD have been conducted in many European and American cities, which revealed that diurnal variations in PSD were linked closely with traffic-related emissions ,24,25,26Meteorological parameters, including temperature, relative humidity (RH), precipitation, wind speed, and planetary boundary layer height, appreciably influence PSD. Paatero  reportedAs mentioned above, numerous studies on PSD were performed in developed countries. Meanwhile, China is also a large source region for anthropogenic aerosols in the world. Measurements of PSD in China have focused mainly on the Beijing-Tianjin-Hebei area, Yangtze River Delta region, Pearl River Delta region, and northwestern China ,43,45; h2, CO, SO2, and O3), BC, and meteorological parameters to explore the key factors causing the durability of high particle number concentrations during winter in Wuhan.This study obtained two years of number concentration data of particles with diameters ranging from 15.1 nm to 661.2 nm from the city of Wuhan since June 2012. The diurnal, monthly, and seasonal variations of PSDs and concentrations were analysed. In addition, the variation characteristics in temperature, RH, atmospheric pressure (AP), precipitation, wind speed, and mixing height and their relationships with particle concentrations in the nucleation mode, Aitken mode, and accumulation mode were investigated in detail. Moreover, a case analysis of a nucleation burst event in winter was conducted using air pollutants  has experienced dense urbanization and industrial activities, with the production of steel from the Wuhan Iron and Steel Corporation ranking fifth and eighth in the world, respectively, in 2013 and 2014 . Figure −1 and 0.3 L·min−1, respectively. A bipolar charger in an electrostatic classifier  was used to charge the particles to a known charge distribution. These charged particles were classified by a differential mobility analyser according to their traverse ability in an electrical field, and a condensation particle counter  was used to count the number concentration of particles in a respective size range. To ensure the quality of data, the volume of n-butyl alcohol and operational parameters, including temperature, voltage, and flow rate, were checked daily, and the valve connector was cleared weekly. Furthermore, the equipment software corrected for the instrumental multi-charged effect and diffusional loss. Excluding missing data and problematic data, 676 integrated days of valid particle number concentrations were recorded from June 2012 to May 2014.Continuous hourly number size distributions of particles between 15.1 nm to 661.2 nm in diameter were measured using a scanning mobility particle sizer  from 1 June 2012 to 31 May 2014. The ambient air was sampled by the SMPS via an approximately 5 feet long and 0.5 inch wide stainless steel tube through the roof of the laboratory, which is about 50 feet above the ground in Wuhan University, with the sheath and sample flows kept stable at 3.0 L·minThe hourly mass concentration of BC was measured using an AethaLometer , based on the estimation of mass concentration of BC that accumulates on a 1.6 cm quartz fibre filter every two minutes. The raw data measured by the AethaLometer was divided into seven channels acquired by wavelengths of 350 nm, 470 nm, 521 nm, 590 nm, 660 nm, 880 nm, and 950 nm. This study only utilized measurements at 880 nm, as this wavelength is considered the standard channel to exclusively estimate BC concentration . As an A2, CO, SO2, and O3, measured at the three national air quality monitoring sites encircling the experimental site, were downloaded from the China National Environmental Monitoring Centre’s website (http://113.108.142.147:20035/-emcpublish/). The continuous monitoring systems for NO2, CO, SO2, and O3 were composed of the sampling unit, analytical device, data collection unit, and calibration instrument. CO was measured adopting the gas filter correlation infrared absorption method and the nondispersive infrared absorption method (API model 300A). O3, SO2, and NO2 were measured via the UV-spectrophotometry, ultraviolet fluorescence (API model 200A), and chemiluminescence  methods, respectively [The hourly mean concentrations of trace gases, comprising NOectively . SimilarpD < 30 nm), Aitken mode (30 nm < pD < 100 nm), accumulation mode (100 nm < pD < 1 μm), and coarse mode (pD > 1 μm) [pD < 30 nm, 30 nm < pD < 100 nm, and 100 nm < pD < 661.2 nm, respectively).PSD is characterized empirically by a multi-lognormal structure, usually divided into four main modes: the nucleation mode ( > 1 μm) . In thisiN (cm−3), 2), and To demonstrate better the characteristics of particle number concentrations and their size distributions, raw data of number size distributions were fitted to a multi-lognormal distribution according to the function :(1)dNd of 0.547, 0.544, and 0.579, respectively. It could be explained that low AP leads to convergence updraft, which promotes the dispersion of particles from near ground to the upper air, resulting in a reduction of particle number concentrations. In contrast, the downdraft caused by high pressure reinforces the downward movement of particles [p < 0.05) of −0.57, −0.515, and −0.563.Similarly, number concentrations of particles in the nucleation, Aitken, and accumulation modes displayed a negative relationship with wind speed, with Pearson coefficients of −0.434, −0.390, and −0.408, respectively, indicating a dilution effect of the wind. However, the Pearson coefficient between wind speed and particle number concentrations in the Aitken mode was not significantly obvious with mosphere . Likewisarticles . Referenarticles  revealedHowever, there rarely existed a correlation between the number concentrations of particles in the three modes and RH. Similarly, the ambiguous relationship between particle number concentrations in the three modes and mixing layer height is shown in The diurnal patterns of particle number concentrations in the different modes averaged over the four seasons are illustrated in As demonstrated via a contour plot of size distributions in number concentration, 4 cm−3 to 2 × 105 cm−3 around 10:00 LT on 3 January and rapidly decreased around 17:00 LT on 5 January, seemingly corresponding to the wind curves shown in 2, NO2, and particles ranging from 30 nm to 200 nm. These additional particles were consistent with the features of long-range transported aerosols described by [2 is one of the primary reactants in photochemical processes, in which atomic oxygen and O3 have been produced [2 oxidizing, a nucleation event emerged with nanoparticles growing into detectable particles by condensation or coagulation, proving that SO2 could be considered a crucial precursor for nucleation events in Wuhan, as it was in Nanjing [From a macro scale, it is obvious that the normalized number concentration of particles in the nucleation mode and partly in the Aitken mode sharply increased from approximately 5 × 10ribed by  that impproduced ,72. Comb Nanjing . The sou2, SO2, and BC, as shown in 2 and CO are produced primarily by high temperature combustion in an urban atmosphere, traffic intensity and industrial activities should be responsible for this type of pollution. Although similar bimodal distributions of trace gases during the traffic rush hours occurred, such similarities were inconspicuous in the contour plot of the size distribution of number concentration in the three modes. A possible explanation for this is that vehicle exhaust emissions were not detectable synchronously under a series of complex chemical reactions or the particle number concentrations of traffic-related emissions were not on the same order of magnitude as the existing pollution. Based on 2, SO2, and BC. Moreover, the O3 concentration, which is considered generally as a proxy for the intensity of photochemical processes, illustrated a unimodal distribution for its diurnal pattern, reaching its peak around 15:00 LT, which is later when compared to usual situations. Furthermore, since NO2 is a depleting substance in photochemical processes, its mass concentration showed a similar profile affected by traffic and industrial emissions simultaneously, but varied in the opposite direction to O3 most of the time. Last but not the least, although particles ranging from 20 nm to 100 nm tended to decrease several times, it never came to fruition because the meteorological conditions were quite stable on 4 January. It could be inferred that the repeated increase in ultrafine particles was attributed to lately generated SO2 via industrial activities, contributing to the “repeated, short-lived” nucleation events, which contrast with the “short-lived” events by [From the diurnal pattern of fine particles, periodic pollution existed in the afternoons and evenings, with increases in mass concentration of CO, NOvents by ,75. In c2, NO2, CO, O3, and BC, were conducted in the city of Wuhan from June 2012 to May 2014. During the experimental period, the average number concentrations of submicron particles in the nucleation (15–30 nm), Aitken (30–100 nm), and accumulation (100–661 nm) modes reached 4923 cm−3, 12,193 cm−3 and 4801 cm−3, respectively. Compared to cities around the world, the particle number concentrations in the three modes in Wuhan were relatively high, directly influenced by the specific level of industry and energy structure.Two-year continuous measurements of particle number concentration and meteorological data and other parameters, including mass concentration of SOPearson coefficients showed that there were distinct differences among the correlation of particle number concentrations in the three modes with precipitation, wind speed, RH, AP, temperature, and mixing layer height. Precipitation, AP, and temperature played a vital role in shaping the number concentration of particles in the three modes, whereas RH and mixing layer height did not appear to have synchronous change with particle number concentrations in Wuhan. Wind speed had a significant negative effect on particle number concentrations in the nucleation and accumulation modes.According to the diurnal variations of particle number concentration in four seasons, particles in the nucleation mode demonstrated a unimodal distribution which reached their peak around noon, with more active nucleation events in summer and autumn. The particles in the Aitken mode showed a triple-peak distribution in spring, summer, and autumn, but a bimodal distribution in winter, possibly caused by a lower rate of new particle formation due to less condensable vapours and solar radiation. The two peaks of Aitken and accumulation mode particles during the morning and evening corresponded to the traffic rush hours, likely due to traffic exhaust emissions.A three-day case analysis was conducted to explore the key factors leading to the durability of high particle concentrations during winter, which contained a complete process comprising a nucleation burst event and a scavenging event. This burst of ultrafine particles resulted from exogenous pollutants, with the thermal inversion layer tremendously impeding the diffusion of particles. These stagnant atmosphere conditions were a hotbed of “repeated, short-lived” nucleation events, combined with dense industrial activities and traffic emissions. The clean northwest wind finally broke the stability of the lower atmosphere, promoting the dispersion of particles with the expansion of the mixing layer.This paper presents a systematic analysis of fine particles in Wuhan, the first such study performed in the central plains of China, and their relationships with climate, meteorological parameters, atmospheric factors, trace gases, and industry levels. To enhance these results, additional factors should be taken into consideration, such as traffic density, detailed industrial emissions, and quantified health effects. In addition, the research into the mechanism of nucleation event and growth ratio about ultrafine particles will be carried out in the next stage. Moreover, this study offers reasonable estimations of particle impacts and provides guidance for policy making and emission control strategies in Chinese cities with heavy industry.


Introduction. Cardiovascular disorders (CVDs) are among the most important diseases in the world and determination of their risk factors is essential for primary and secondary prevention. This study aimed to evaluate these risk factors in workers of tile and ceramic industry, a main industry in Yazd. Materials and Methods. In a cross-sectional study, 1075 tile and ceramic workers were selected by simple sampling method. BMI, blood pressure, FBS, and lipid profile were measured and compared to international standards. Results. 731 individuals (68%) had at least one risk factor, and 52%, 12%, 3%, and 0.7% had one, two, three, and four risk factors, respectively. The most common risk factor was abnormal BMI (49.6%); low HDL (48.4%) and high TG (14.1%) were in the second and third orders. Conclusion. This study showed a relatively high prevalence for CVD risk factors among tile and ceramic workers. Low HDL, high TG, and overweight were the most frequent risk factors in this population. Cardiovascular disorders (CVDs) are among the most important and frequent chronic and noncommunicable diseases in the world and contribute to 30% of mortalities throughout the world . AccordiAtherosclerosis and its related factors (risk factors) cause coronary artery disease . These rPhysical activity is a predictor of CVDs and their mortality . NowadayIt is estimated that worldwide prevalence of hypertension is about one billion and it causes approximately 7.1 million deaths per year . AccordiOther occupational factors which can increase the risk of CVDs include high workload, shift work, noise, and psychosocial factors , 12. SomSome studies have assessed the cardiovascular risk factors among occupational groups. In a study in China, the most prevalent cardiovascular risk factors in steel workers included hypertension, smoking, overweight, and hypercholesterolemia . In anotEvaluation of adults' cardiovascular risk factors is an important issue to reduce the frequency of disorders and their complications. Employees as a large and active population in a society are apt to the risk factors of CVDs some of which may be preventable. So in this study we aimed to evaluate the cardiovascular risk factors of workers of tile and ceramic industry which is the main industry in Yazd. We couldnot find another study conducted on tile and ceramic workers regarding cardiovascular risk factors.In a cross-sectional study from March 2011 till March 2012, 1200 tile and ceramic workers were selected by simple sampling method. There were only 48 females who were excluded from the study due to low sample size. Among males, 77 subjects did not continue the study, so at last the results of physical and lab tests of 1075 workers were evaluated. Workers were selected from different parts of the factory, including grinding, ball mill, forming, glazing and printing, glaze-making, firing, packing, office, service, laboratory, maintenance, forklift driving, and warehouse.Medical history taking and physical examination were performed by three occupational physicians who were similarly trained for this project. Demographic data  were recorded for each participant.Individual or family history of CVDs was asked by the physicians. Physical examination included measuring height, weight, and blood pressure. Height was measured in standing neutral position without shoes. Weight was measured by a digital scale  with subjects wearing home clothing without shoes. Waist circumference was measured at 2-3 cm above the umbilicus. Hip circumference was defined as the greatest diameter between the waist and knee . BMI wasBMI was categorized according to WHO classification, that is, BMI < 18.5: underweight, 18.5 ≤ BMI ≤ 24.9: normal, BMI ≥ 25: overweight, 25 < BMI ≤ 29.9: preobesity, and BMI ≥ 30: obesity . AbdominBlood pressure was classified according to the guidelines of European Society of Hypertension and European Society of Cardiology, that is, SBP < 120 and DBP < 80: optimal, 120 ≤ SBP ≤ 129 and 80 ≤ DBP ≤ 84: normal, 130 ≤ SBP ≤ 139 or 85 ≤ DBP ≤ 89: high normal, and SBP ≥ 140 or DBP ≥ 90: hypertension [Blood sampling was performed in the morning with at least 12-hour fasting to measure fasting blood sugar (FBS), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), and low density lipoprotein (LDL). Blood parameters were analyzed by an autoanalyzer (Hitachi 902). All laboratory tests were performed in a single laboratory.FBS level was classified based on the recommendations of the American Diabetes Association, that is, FBS < 100: desirable, 100 ≤ FBS < 125: borderline, and FBS ≥ 126: diabetes .Blood lipids level was categorized according to the National Cholesterol Education Program (NCEP), that is, TG < 150: normal, 150 ≤ TG < 199: borderline, TG ≥ 200: increased. LDL < 100: optimal, 100 ≤ LDL ≤ 129: desirable, 130 ≤ LDL ≤ 159: borderline, and LDL ≥ 160: increased.HDL ≥ 40: desirable, HDL < 40: decreased .t-test and chi square test. An informed consent was obtained from each participant (in Persian). Level of significance was considered to be 0.05. This study was approved by the Ethics Committee of Shahid Sadoughi University of Medical Sciences.Data were analyzed by SPSS (ver. 19) using student's Study subjects included 1075 tile and ceramic workers. All participants were males with mean age of 31.38 ± 6.77 years. 78% of them were married. Thirty-four participants (3.1%) had completed elementary education and 123 ones (11.5%) had completed guidance school. In addition, 871 (4.4%) and 48 (81%) participants were graduated from high school and university, respectively.Among all subjects, 731 individuals (68%) had at least one risk factor, and 52%, 12%, 3%, and 0.7% had one, two, three, and four risk factors, respectively. Tables Cardiovascular diseases are among the most important causes of morbidity and mortality in different populations. Sedentary life style is one of the main risk factors of these diseases, so in occupational settings with low activity we expect to find a lot of risk factors, but in industrial settings in which workers are active during work shift, it seems that the frequency of CVD risk factors is low.The main CVD risk factors include age, race, gender, smoking, hypercholesterolemia, type II diabetes, hypertension, obesity and overweight, sedentary life style, and stress . Some ofIn this study, we assessed the frequency of different CVD risk factors among tile and ceramic workers considering different job titles in this industry. This study identified a relatively active and young population with a high prevalence of cardiovascular risk factors.Although overweight and obesity were not included in the general CVD risk prediction model of the Framingham Heart Study , the impIn our study, overweight was observed in about half of the subjects and obesity in about 15% of them, which is very high if we consider that we have evaluated a relatively young and active population, because they were industrial workers who are completely active at least for 8 hours in their working shift, although this frequency was lower than those of Yazd urban population with 16.5% obesity , BraziliThe mean BMI in this study was similar to that in Tehran urban population (25.7%), but the frequency of overweight was lower in that study (40%) ; althougHypertension as one of the other important risk factors of CVDs was lower in our study compared to 27.59% in Yazd adult population  and 20.4Frequency of abnormal FBS in this study was lower than those of Yazd urban population , Tehran The frequency of smokers in this study was lower than those of Yazd urban population , PrabhakMean TG level was similar to those of industrial workers in India  and bus Hypertriglyceridemia in this study was lower than those of Indian industrial workers , IsfahanOther risk factors such as increased LDL and cholesterol were also different in this study from other studies on different jobs , 29, 31.The most frequent risk factor in this study was abnormal BMI; Low HDL and increased TG were in the 2nd and 3rd orders. Overweight was most commonly observed in warehouse workers which can be explained by their lower level of activity compared to other workers. Overweight in office workers, who have sedentary work as well, was lower than in warehouse workers, which is probably due to their higher social level and different nutrition style.This study showed a relatively high prevalence for CVD risk factors among tile and ceramic workers. Low HDL, high TG, and overweight were the most frequent risk factors in this population.


The present study investigated the relationship between Chinese reading skills and metalinguistic awareness skills such as phonological, morphological, and orthographic awareness for 101 Preschool, 94 Grade-1, 98 Grade-2, and 98 Grade-3 children from two primary schools in Mainland China. The aim of the study was to examine how each of these metalinguistic awareness skills would exert their influence on the success of reading in Chinese with age. The results showed that all three metalinguistic awareness skills significantly predicted reading success. It further revealed that orthographic awareness played a dominant role in the early stages of reading acquisition, and its influence decreased with age, while the opposite was true for the contribution of morphological awareness. The results were in stark contrast with studies in English, where phonological awareness is typically shown as the single most potent metalinguistic awareness factor in literacy acquisition. In order to account for the current data, a three-stage model of reading acquisition in Chinese is discussed. Children typically develop literacy skills in literate societies with appropriate education. However, there are some children who fail to acquire reading proficiency, who tend to have associated behavioral, academic, and psychological difficulties Extensive research into the cognitive processes involved in reading has shown that in alphabetic languages phonological awareness plays a dominant role e.g., . A similMorphological awareness is defined as children's conscious awareness of the morphemic structure of words and the meaning of morphemes, which are the smallest meaningful linguistic units An increasing body of research has shown converging evidence for the importance of morphological awareness in reading Chinese. McBride-Chang, Shu, Zhou, Wat and Wagner nd grade children, to obtain the pronunciations of consistent/regular characters Orthographic awareness refers to the reading processes involved in forming, storing, and accessing the orthographic representations of a language The first aim of the current study was to determine which metalinguistic awareness skills such as orthographic, phonological and morphological awareness exert a unique influence on the acquisition of literacy in Chinese. Do orthographic awareness skills play a more important role than the other two metalinguistic awareness skills, or do morphological awareness or phonological awareness skills exert a stronger influence in reading Chinese? For significant correlations among the three metalinguistic awareness skills, it is important to control any two of them to observe the unique contribution of the third metalinguistic awareness skill. Further, rapid automatized naming, phonological memory and nonverbal intelligence as control variables should also be measured, since they also have been shown as influential predictors in the acquisition of reading skills in Chinese The second aim was to determine if these metalinguistic awareness skills would exert their influence at different developmental stages in the acquisition of literacy in Chinese. Models of English word-reading development along with empirical evidence supported the idea that the independence and importance of the contribution of phonological, morphological, and orthographic awareness skills to word reading varied with age e.g., . Ho et a411 children from two primary schools in Shijiazhuang, the capital of Hebei province participated in the study. 20 children were excluded from the analyses due to their incomplete data. The remaining participants consisted of 101 preschool children , 94 Grade-1 children , 98 Grade-2 children , and 98 Grade-3 children . All children were Mandarin speakers. None of the children were reported to have a history of language or speech or hearing problems by their teachers. Written consent was obtained from the children's parents and their teachers. The research project was approved by the Research Ethics Committee of the Institute of Psychology, Chinese Academy of Sciences.The materials consisted of a battery of tests covering phonological awareness, orthographic awareness, morphological awareness, naming, phonological memory and character recognition. Each test is described in detail below. All the tasks were administrated within two sessions, and each session took about 40 minutes.Oddity Detection task and Deletion task were used to measure (i) tone awareness, (ii) onset-rime awareness and (iii) phonemic awareness. The tasks that measure different levels of phonological awareness were included in order to prevent floor and ceiling effects when the same measures were given across a wide range of children's ages following Gottardo, Yan, Siegel, & Wade-Woolley In the Oddity Detection test, the children were asked to find the odd syllable within three stimuli presented orally twice by the experimenter. For example, /hao3/ is the odd one in the set /tan2/, /gan4/, /hao3/.There were two practice and 14 experimental trials. This test showed reliability of 0.60 . In the Deletion test, the children were asked to say a syllable aloud after deleting a specified phoneme, onset or rime of a character  presented orally by the experimenter. For example, say /miao2/ without the /i/ sound. Prior to 14 experimental trials, two practice trials were given. The stimulus items were based on the phonological tests by Liu, Liu, & Zhang Sentence A and Sentence B were orally presented to the participants, indicating a certain morphological relationship, such as, “Early in the morning we can see the sun rising. This is called a sunrise.” Then Sentence C was presented to create a new scenario, and children were asked to produce a new word or concept to complete Sentence D based on the morphological relationship. For example, Sentence C and D for this trial would be “At night, we might also see the moon rising. What could we call this?” The correct answer is moonrise. There were 15 stimulus items, six of which were adapted from McBride-Chang et al. Morphological awareness skills were measured by a morphological construction test and a morpheme judgment test. The Chinese morphological construction test was similar to a sentence analogy test (see below), which was often used to measure morphological awareness skills in English Morpheme judgment test was used to investigate children's ability to understand the different meanings of one character as a constructed morpheme in different words. For example, “There were two practice trials prior to 30 experimental trials, half of which were from Shu et al. Similar to the previous studies Print knowledge of Chinese characters refers to the rudimentary knowledge of Chinese orthographic regularity, that is, understanding of the rudimentary orthographic regularities/conventions of printed Chinese characters. The test consisted of 45 two-alternative stimulus items, in which non-characters violated certain print conventions of Chinese characters  (Contemporary Chinese Frequency Dictionary) by the stratified random sampling method used by Chen Character Recognition test were used to measure reading ability. 110 Chinese characters were used as stimuli, 100 of which were selected from In addition, rapid automatized naming, phonological memory and nonverbal intelligence were measured as control variables. Naming performance was measured by a continuous number-naming task similar to that used by Denckla and Rudel p<.01 for Preschool, and r = .48, p<.01 for Grade-1 children, but for Grade-2 children, the skills were significantly related to the Oddity Test, r = .42, p<.01, and for Grade-3 children, the character recognition skills were significantly related to the performance in Morphological Construction Test, r = .37, p<.01.Thus, a significant and positive relationship between the character recognition skills and orthographic awareness skills was found in younger (Preschool and Grade-1) children, but not in the older (Grade-2 and Grade-3) children. Conversely, a significant positive relationship between the character recognition skills and Pseudo-word Repetition was found in older (Grade-2 and Grade-3) children, r = .24, p<.05 for Grade-two, and r = .31, p<.01 for Grade-3 children, but not in the younger children.The results revealed the following: for Preschool and First-grade children, character recognition skills were significantly related to Lexical Decisions, r = .63, Finally, in order to directly compare the relative contributions of Phonological, Morphological, and Orthographic Awareness, hierarchical multiple regressions were conducted with the following variables being controlled: Nonverbal Intelligence, Naming and Phonological Memory. The data from the two Phonological Awareness tests, the two Morphological Awareness tests, and the two Orthographic awareness tests were entered in the equations separately in the final step. The results of the analyses are shown in p<.001; 4.7%, F  = 3.70, p<. 05; and 20.7%, F  = 22.21, p<.001 of the variance of Character Reading, respectively.For Preschool children, Phonological Awareness, Morphological Awareness and Orthographic awareness accounted for 11.7%, F  = 10.44, p<.01 of the variance.For Grade-1 children, neither Phonological Awareness nor Morphological Awareness accounted for a significant portion of the variance of Character Reading. Orthographic awareness was the only significant predictor for Character Recognition in these children, which accounted for 8.6%, F  = 5.80, p<. 01 and 6.0%, F  = 4.00, p<.05 of the variance respectively in Character Recognition. For Grade-3 children, Morphological Awareness was the only significant predictor of the variance for Character Recognition, which accounted for 7.3%, F  = 4.48, p<.05.For Grade-2 children, two significant predictors were Phonological Awareness and Morphological Awareness, which accounted for 10.4%, F  = 7.47, The present study revealed that reading development through preschool/kindergarten to third-grade Chinese children was influenced by a comprehensive set of cognitive skills. The relationship between reading ability and these different metalinguistic skills appears to change with age. In the following section each of the important cognitive skills identified in the current study in relation to the literacy acquisition/development in Chinese are discussed in turn.As with the previous studies e.g., , the curMoreover, the current results showed that phonological awareness significantly predicts the character recognition at preschool and Grade-2, but not at Grade-1 and Grade-3.  Similarly, Siok and Fletcher From The unique contribution of Orthographic Awareness independent of the other factors was identified in this study. The importance of orthographic processing skills in reading in Chinese has been discussed by other researchers e.g., . HoweverAs shown in However, the contribution of orthographic knowledge to reading acquisition appeared to become insignificant for older children . This developmental pattern in Chinese is very different from that of English. In English, the acquisition of the orthographic knowledge requires sufficient print exposure Caution is needed in drawing any conclusions about the relationship between orthographic awareness and the reading ability of children older than Grade-1 because of the ceiling effect . Despite the fact that Ho and his colleagues As described earlier, Ho et al. One of the most logical directions for future research lies in a longitudinal investigation to consolidate the findings obtained from the present cross-sectional study. Although it is acknowledged that the interpretations offered in this study may still be tentative, they provide a general profile of Chinese reading acquisition in young children. Another line of new research lies in the inclusion of other variables such as size of vocabulary Click here for additional data file.Table S2Three Types of Non-character in the Lexical Decision Task.(DOC)Click here for additional data file.


Long-term care residents are susceptible to constipation and one-half to three quarter of older nursing home residents receive laxatives regularly.The purpose of this study was to evaluate the factors related to abnormal bowel function and explore the effectiveness of laxative treatment among the elderly residents of a nursing home.A total of 110 residents older than 65 years old was enrolled in this study. The following variables were gathered: age, gender, body mass index (BMI), length of stay, daily fluid intake, type of food, functional level, cognitive ability, physical therapy status, somatic and psychiatric diseases, number of medications, and medication use. The use and dosage of laxatives were recorded by means of Anatomical Therapeutic Chemical (ATC) classification system. Normal bowel function was defined as defecation frequency from three defecations per day to three defecations per week and stool consistency score of three to five on Bristol Stool Form Scale. A comparison between groups with normal and abnormal bowel function was drawn.Low BMI, increased fluid intake, liquid food intake, poor functional level, poor cognition, and a history of stroke were significantly associated with altered bowel function (P < 0.05). The most frequently used laxatives were glycerol, senna glycoside, and magnesium oxide. There were significant differences in laxative regimens between residents with normal and altered bowel function; those with altered bowel function tended to take more laxatives than those with normal bowel function.This study suggested that treatment of constipation in the nursing home was unsatisfactory. To improve treatment outcomes in those susceptible to altered bowel function, a coordinated approach with involvement of physicians, nursing staff, and other professionals including dieticians and pharmacists seems necessary. Constipation is a prevalent problem among elderly population. Complaints of constipation and its prevalence increase with age, particularly after the age of 65 , 2. The The purpose of this study was to evaluate the factors related to the abnormal bowel function and explore the effectiveness of laxative treatment among the elderly of a nursing home. The clinical trial was conducted in a certified, professionally run nursing home, which is an affiliated facility of a regional hospital in Hsinchu County, Taiwan, from January to June in 2012. In addition to those from Hsinchu area, many of the residents come from the nearby counties or cities. A physician examined and evaluated all the residents admitted to the nursing home for past medical history, current physical and functional conditions, and prescription of the routine medications. Daily prescribed medication of the residents were formally recorded. From January to June in 2012, residents older than 65 years old in the facility were evaluated. Those who had stayed at the nursing home for more than six months were included in this study. Those who left the nursing home for another institution or returned home were excluded from the study. We further excluded those residents with known organic gastrointestinal diseases such as cancer, inflammatory bowel disease, and intestinal resection. Finally, A total of 110 residents were enrolled in this study. The following variables were gathered: age, gender, body mass index (BMI), length of stay, daily fluid intake, type of food , functional level scored by Barthel Index , cognitiThe use and dosage of laxatives was recorded by means of the Anatomical Therapeutic Chemical (ATC) Classification System. Groups of laxatives were defined at ATC-level 5: osmotically acting laxatives (A06AD) such as magnesium oxide and lactulose; contact laxatives (A06AB) such as bisacodyl and senna glycosides; bulk laxatives (A06AC) such as sterculia; enemas (A06AG) such as glycerol and sorbitol; and softeners/emollients (A06AA) such as liquid paraffin. The dosage of each laxative was graded as on demand, regular use of standard dose, and regular use of high dose. High dose was defined as bisacodyl > 10 mg/day; senna glycosides > 24 mg/day; sterculia > 2 package (14GM) per day; lactulose > 30 mL/day; and magnesium oxide > 1000 mg/day .Defecation frequency (number of stools per day), stool consistency , straining, sensation of incomplete evacuation, sensation of anorectal obstruction, and manual maneuvers to facilitate bowel movements were recorded. Bristol Stool Form Scale can be used to monitor change in intestinal function. The seven types of stool are as follows: type 1, separate hard lumps, like nuts; type 2, sausage-shaped, but lumpy; type 3, like a sausage but with cracks on its surface; type 4, like a sausage or snake, smooth and soft; type 5, soft blobs with clear cut edges; type 6, fluffy pieces with ragged edges, a mushy stool; and type 7, watery, no solid pieces . In our This study was performed in compliance with the guidelines for research involving humans and was approved by the institutional review board of the National Taiwan University Hospital Hsin-chu Branch.Comparisons between groups were performed with the Independent t-test, the exact chi-square test, and Fisher’s exact test using Statistical Package for the Social Sciences . In all tests, P-values < 0.05 indicated a statistically significant difference.The study included 110 residents who were older than 65 years and had been continuously staying in the nursing home for more than 6 months. The mean age of the participants was 80.7 ± 7.6 years, ranging from 65 to 99 years. Participants included 55 males and 55 females. The average duration of nursing home stay for all subjects was 40.7 ± 25.7 months, ranging from six to 117 months. The number of diseases of all subjects was categorized and listed as nil in 21 , one in 49 , two in 30 , and three in 10 participants . There was no significant difference between the group 1 and group 2 in terms of the number of diseases. The number of medications of all subjects was categorized and listed as 0 to 4 in 42 , 5 to 8 in 49 , and more than 8 in 19 participants . There was no significant difference between the group 1 and 2 in the number of medications. There were 18 (16.4%) participants having no record of laxative use or only using an enema on demand; a total of 92 (83.6%) residents in our study used laxatives regularly. Only nine (8.2%) participants did not take laxatives during the investigation period and all of them had normal bowel function. In other words, 101 (91.8%) participants ever took laxatives during the investigation period. Senna glycoside was the most commonly and regularly used laxative in this study , and glycerol enema was the most frequently used laxative on demand . Senna glycoside generally induces evacuation eight to 12 hours following administration and can be taken at bedtime. In addition to the convenience, senna glycoside is a cheap and safe agent for use in the elderly . Glycero


Background. Acupuncture is applied to treat numerous diseases in pediatric patients. Few reports have been published on the depth to which it is safe to insert needle acupoints in pediatric patients. We evaluated the depths to which acupuncture needles can be inserted safely in chest acupoints in pediatric patients and the variations in safe depth according to sex, age, body weight, and body mass index (BMI). Methods. We retrospectively studied computed tomography (CT) images of pediatric patients aged 4 to 18 years who had undergone chest CT at China Medical University Hospital from December 2004 to May 2013. The safe depth of chest acupoints was directly measured from the CT images. The relationships between the safe depth of these acupoints and sex, age, body weight, and BMI were analyzed. Results. The results demonstrated significant differences in depth among boys and girls at KI25 (kidney meridian), ST16 (stomach meridian), ST18, SP17 (spleen meridian), SP19, SP20, PC1 (pericardium meridian), LU2 (lung meridian), and GB22 . Safe depth significantly differed among the age groups (P < 0.001), weight groups (P < 0.05), and BMI groups (P < 0.05). Conclusion. Physicians should focus on large variations in needle depth during acupuncture for achieving optimal therapeutic effect and preventing complications. Acupuncture has been used for thousands of years based on the theory of meridians and is crucial component of traditional Chinese medicine. Acupuncture has been applied to numerous diseases and has been found to be efficacious in treating chronic pain, postoperative nausea and vomiting, postoperative pain, chemotherapy-induced nausea, acute pain including dental pain, headache, hypertension, and chronic obstructive pulmonary disease (COPD), and seasonal allergic rhinitis , 2. AcupAcupuncture may lead to severe complications, such as the transmission of diseases, needle fragments left in the body, nerve damage, pneumothorax, pneumoperitoneum, organ puncture, cardiac tamponade, and osteomyelitis . PneumotTraditionally, the depth of needling at acupoints was measured by cun and has variations in ancient book, which has no difference between people of different body size. Owing to the fact that there is a lack of consistency in determining the depth of insertion, the depth of needle insertion was usually judged by clinical experience of acupuncture practitioners and the response from the patients. In children, it may be difficult to elicit a proper response to the needle insertion. Thus, it is critical to establish the data of acupoint depth for safe needling, specific for children.A few surveys have been done by different measuring tools to investigate safe needle depth . These sMost previous studies have involved small sample sizes and adult groups. In pediatric groups, changes in muscle mass and fat volume and their distribution, particularly during the puberty stage, must be considered because of the influence on safe needle depth, which is unnecessary in adult groups. Scant studies on safe needle depth have been conducted in pediatrics. Only 2 previous studies, both conducted by Chen et al., have included pediatric groups; they measured the safe depth and therapeutic depth of abdomen acupoints according to CT scans , 16. In All patients aged 4 to 18 years who had undergone a chest CT between December 2004 and May 2013 at China Medical University Hospital (CMUH) were selected. These patients had undergone CT scanning to evaluate acute chest condition, such as acute accidental injuries, pneumonia, pneumothorax, and cardiac diseases. Patients with chronic oncology diseases, chest-wall deformity, or chest-wall trauma were excluded because of the possible effect on the thickness of subcutaneous tissues or muscles of the chest-wall. Patients diagnosed with pneumothorax were measured on the contralateral side.2 (m2). Patients were divided into 5 groups according to age: 4 to 6 (48–83), 7 to 9 (84–119), 10 to 12 (120–155), 13 to 15 (156–191), and 16 to 18 (192–216) years (mo). Patients were also divided into 4 groups based on weight: below the third percentile, between the third and 85th percentiles, between the 85th and 97th percentiles, and at or above the 97th percentile, according to the new growth charts for Taiwanese children and adolescents [The age, sex, and body weight of each patient were obtained from chart records. BMI was measured according to weight (kg)/heightlescents . Patientlescents , 18. TheThe acupoints were located according to a classical Chinese acupuncture technique, known as Tong Shen Cun (cun). The chest transverse Tong Shen Cun is one-eighth of the distance between the 2 nipples. The chest vertical Tong Shen Cun is 8 cuns of distance between CV22 to CV17. CV22 is located at the central point of 2 clavicles and the suprasternal notch. CV17 is located at the central point of the 2 nipples and on the sternum. The safe depths of acupoints in the conception vessel (CV) meridian (Renmai) were defined as the distance from the skin surface of the acupoint to the sternum, except CV22. The safe depth of CV22 was the distance from the skin to the trachea, and the safe depths of other acupoints over the chest were defined as the distance from the skin surface of acupoints to the pleura.In total, 28 acupoints located at the conception vessel meridian (CV) (Renmai), kidney meridian (KI), stomach meridian (ST), spleen meridian (SP), gallbladder meridian (GB), lung meridian (LU), and pericardium meridian (PC) were measured. Patients diagnosed with pneumothorax were measured on the healthy side. From CV21 to CV16, the interval of each acupoint was 1.6 cuns. The kidney meridian, stomach meridian, and spleen meridian were 2, 4, and 6 cuns from the CV meridian. GB23 and GB22 were 7 and 8 cuns from CV17. LU2 was 6 cuns from CV21, and LU1 was the same distance from CV20. PC1 was 5 cuns from CV17.The CT machines used at CMUH were the Light Speed16 CT Scanner , Optima Speed CT Scanner , and Optima CT660 . All chest CT images were captured in the transverse plane. The section thickness between each image was 5 mm. The safe depth was measured according to the CT image displayed on a Picture Archiving and Communication System (PACS) monitor .The safe depths of acupoints among different groups of weight, age, and BMI were analyzed using one-way analysis of variance (ANOVA) or Brown-Forsythe test. Brown-Forsythe test was used when the variances were heterogeneous. Two-way ANOVA was then used to determine the effects considering the sex in the analysis of the other variables.P < 0.05 was considered significant.The generalized linear model was used to analyze whether sex, age, weight, and BMI are relevant factors in determining the safe depth of acupoints. Statistical analyses were performed using the SPSS software package, version 18.0  and From December 2004 to May 2013, 319 patients (205 boys and 114 girls) ranging from 4 to 18 years of age were included in this study. All participants were Taiwanese. Among them, only 318 of those had weight data and 286 of those had height data to get BMI. The mean and standard deviation of the safe depth for boys and girls for 28 chest acupoints are shown in P < 0.001) (Safe depth significantly differed among age groups (< 0.001) . Age wasP < 0.05) (Safe depth significantly differed between weight groups ( < 0.05)  and signSafe depth also significantly differed between BMI groups . An incrWe also applied generalized linear regression modeling to analyze whether age, sex, weight, and BMI are relevant factors in determining the safe depth of acupoints. The results revealed that sex and weight were the most crucial factors independent of age and BMI in determining the safe depth of chest acupoints (data not shown).This study used retrospective data and chest CT images from 2004 to 2013 in CMUH to estimate the variations in safe depth according to sex, age, weight, and BMI in children aged 4 to 18 years. We observed a significant difference in safe depth at KI25, ST16, ST18, SP17, SP19, SP20, PC1, LU2, and GB22 between both sexes, which significantly differed among the age groups, weight groups, and BMI groups.In this study, the safe depth at KI25, ST16, ST18, SP17, SP19, SP20, and PC1 acupoints in girls was deeper than it was in boys. These acupoints were significantly deeper in girls around the breast. Thelarche in girls typically begins between the ages of 8 and 13 years, with an average onset age of 10.3 years . This seP < 0.001). However, safe depth significantly correlated with an increase in age only in KI23, ST17, LU2, SP17, and GB22. At another 23 chest acupoints, safe depth did not increase with age in the 10-to-12-year-old and older groups. At each acupoint, depth increased significantly with age only before 12 years of age. During puberty, boys gain greater amounts of fat-free mass and skeletal mass, whereas girls acquire significantly more fat mass [Safe depth significantly differed among age groups (fat mass . This maSafe depth significantly increased with weight, except at CV21 and CV22 acupoints. A thicker fat or muscle tissue layer in heavier children may explain this observation. This result is compatible with an adult study in which the safe depths in chest acupoints were related to body size [The BMI characterizes the relative proportion between the child's weight and height. The BMI is a valid predictor of adiposity and is therefore the optimal clinical standard for defining obesity in adults and in children older than 2 years of age . BecauseGeneralized linear regression analysis revealed that sex and weight were relevant factors independent of age and BMI in determining the safe depth of chest acupoints. Previous studies had observed that BMI does not predict body fat very well in low to normal BMI children , 27. BesThis study is the first to investigate safe depth of chest acupoints in pediatric groups. We also studied factors such as sex, age, weight, and BMI that influence safe depth. However, our study has limitations, which include the retrospective design and the small sample size in the overweight group. In addition, our study participants were patients, not healthy children; however, we excluded patients with diseases that might affect the thickness of subcutaneous tissues or muscles of the chest-wall. Finally, because a single study may cause regional bias and all participants were Taiwanese children, international studies with a larger distribution should be conducted.This study demonstrated that safe depths of chest acupoints around the breast were significantly deeper in girls. Safe depth increased significantly with age before 12 years of age. An increase in the weight and BMI was significantly correlated with the increase in safe depth. Thus, children with heavier weight would have deeper safe needle depth over chest acupoints. Physicians should focus on large variations of needle depth when performing acupuncture for achieving optimal therapeutic effects and for preventing complications. Future works about safe needle depth at acupoints over other body parts in children should be conducted and a formula considering weight or BMI index may be created for safe acupuncture.


The topsoil around Beijing metropolis, China, is experiencing impacts of rapid urbanization, intensive farming, and extensive industrial emissions. We analyzed the concentrations of Cu, Ni, Pb, Zn, Cd, and Cr from 87 topsoil samples in the pre-rainy season and 115 samples in the post-rainy season. These samples were attributed to nine land use types: forest, grass, shrub, orchard, wheat, cotton, spring maize, summer maize, and mixed farmland. The pollution index (PI) of heavy metals was calculated from the measured and background concentrations. The ecological risk index (RI) was assessed based on the PI values and toxic-response parameters. The results showed that the mean PI values of Pb, Cr, and Cd were > 1 while those of Cu, Ni, and Zn were < 1. All the samples had low ecological risk for Cu, Ni, Pb, Zn, and Cr while only 15.35% of samples had low ecological risk for Cd. Atmospheric transport rather than land use factors best explained the seasonal variations in heavy metal concentrations and the impact of atmospheric transport on heavy metal concentrations varied according to the heavy metal types. The concentrations of Cu, Cd, and Cr decreased from the pre- to post-rainy season, while those of Ni, Pb, and Zn increased during this period. Future research should be focused on the underlying atmospheric processes that lead to these spatial and seasonal variations in heavy metals. The policymaking on environmental management should pay close attention to potential ecological risks of Cd as well as identifying the transport pathways of different heavy metals. Excessive accumulation of heavy metals is not only toxic to animals and plants , 2, but The concentration of heavy metals in the topsoil is naturally determined by parent materials and subsequent pedogenic processes. However, the anthropogenic input of heavy metals is becoming the main source of heavy metals in the soils of many regions –7. Agroc2, including Beijing, Tianjin, and parts of the provinces of Hebei, Shandong, and Shanxi. The total population of the region in 2010 was approximately 146 million, accounting for 10.7% of the nation’s entire population. As one of China’s most important food bases, the HRB produced 10% of the total grain yield, despite only occupying 3.3% of the nation’s overall land area [In developing countries such as China, there are many problems associated with heavy metal pollution due to the rapid urbanization and intensive anthropogenic activities , 20. Theand area . The HRBThe long-term impairment from heavy metal pollution in this region is regarded as a serious danger to mankind, resulting in the “cancer villages” around Beijing reported by China Central Television (CCTV) in 2013 , 23. TheThe objective of this study is to: investigate the pollution levels and ecological risks associated with Cu, Ni, Pb, Zn, Cd, and Cr around Beijing metropolis; and assess the impact of land use types and crop rotations on the variations of heavy metals during the pre- and post- rainy seasons.As one of the largest watersheds in China, the HRB is located at 35°3′23″N—42°43′48″N and 111°58′4″E—119°49′15″E. The HRB region has a warm-temperature climate  with a mWe selected sampling sites selected based on the representativeness of the land use types and topographic features as well as the ease of access. No specific permissions were required for these locations and activities in the field sampling and we confirmed that the field studies did not involve any threat to endangered or protected species.A total of 202 topsoil samples (0~20 cm) were collected in May and August of 2010, representing the pre- and post-rainy season. The specific sampling sites were selected away from the main roads in an attempt to reduce the vehicle impact. Five samples were taken around the center of each site after removing the plant residues using a spade. The samples were mixed thoroughly to select 1000 g of soil as the representative sample for the site and stored in a plastic bag. We recorded the site features including coordinates, altitude, land use type, and topography. In the pre-rainy season, 87 samples were collected from different land use types including forest (21 samples), shrub (9), grass (8), spring maize (12), orchard (12), wheat (20), and mixed farmland (5). In the post-rainy season, 115 samples were selected from the forest (25), shrub (15), grass (9), summer maize (42), orchard (10), cotton (7), and mixed farmland (7). The mixed farmland was used for planting quick-growing crops in mixed patterns, such as beans, potatoes, and peanuts. Wheat was the staple crop and was rotated in one year by the summer maize or the cotton. The spring maize was a single-season crop in the northern China. We examined eight combinations of land use comparison and crop rotations from the pre- to post-rainy season as follows: forest—forest, grass—grass, shrub—shrub, orchard—orchard, mixed farmland—mixed farmland, spring maize—summer maize, wheat—summer maize, and wheat—cotton. The samples covered most of topographic features and land uses around the Beijing metropolis . Road da4-HNO3 (5:5:3) was added. The samples were heated on a hot plate at 200°C and kept at a slight boiling state until the solid residue disappeared [The soil samples were first air-dried, crushed, and sieved through a 1 mm mesh at room temperature. The soil pH was determined using a standard pH electrode based on 10 g dried soil in 25 ml distilled water. The sieved samples were passed through a 0.15 mm nylon sieve for chemical analysis using Method 3050B (USEPA 1996) , 27, 34.appeared . The Cd  is the measured concentration (mg/kg) of heavy metal i, and i in each province of China. A PI value of heavy metals > 1 indicates the accumulation and potential pollution of heavy metals in the topsoil.The pollution index (PI) associated with heavy metals can be assessed using the measured and background concentrations , 28, 36.Ti is the toxic-response parameter  [Ecological risk index (RI) was used to assess the potential harm caused by heavy metals in the topsoil . The RI  Zn = 1) , 36. Thep < 0.05). All statistical analyses were conducted using SPSS 19.0 .Pearson correlation coefficients were calculated to quantify the relationship among the physical properties and heavy metal concentrations. Analysis of variance was used to test difference in PI between different land use types and different seasons. This analysis indicates that the PI difference is reliable when the significance test is true  were located at high elevations while cultivated land uses  were located at low elevations .Results of Pearson correlation analysis showed that the elevation was significantly correlated with the chemical properties of sampling sites . The soiThe measured concentrations of heavy metals were compared with the background concentrations and the China environmental quality standard for soils (EQSS) . The EQSThe mean PI of all samples increased in the following order: Ni (0.718) < Zn (0.859) < Cu (0.977) < Pb (1.132) < Cr (1.219) < Cd (2.260). The proportion of sampling sites with a PI > 1 was 9.90%, 25.74%, 35.15%, 61.88%, 64.85%, and 93.07% for Ni, Zn, Cu, Pb, Cr, and Cd, respectively. The RI values of Cu, Ni, Zn, Pb, and Cr were less than 40 for all samples which indicated low ecological risks. However, the RI values of Cd indicated high ecological risks in the region. There were 15.35% of total samples with a low ecological risk, 60.4% of total samples with a moderate ecological risk, 22.28% of total samples with a severe ecological risk, and 1.98% of total samples with a serious ecological risk.The heavy metal concentrations around the Beijing metropolis have been investigated in many studies . Generalp value) of PI difference was calculated to assess the reliability of seasonal variations (p < 0.05). However, the results indicated that seasonal variations varied according to different types of heavy metals. Specifically, most PI values for Cu, Ni, and Zn were < 1, both in the pre- and post-rainy seasons. In contrast, Cd had the highest PI values during this period. The PI values for Cr were > 1 in the pre-rainy season only, while they were > 1 for Pb in the post-rainy season. Overall, the mean PI values of Cu, Cd, and Cr decreased from the pre- to post-rainy season, which was opposite to the trend of the PI values of Ni, Pb, and Zn.We compared the mean PI of heavy metals between the pre- and post- rainy seasons. The statistical test  of the PI difference between each pair of land use types (p < 0.05). This result indicated that the impact of land use was not significant compared with the impact of precipitation.se types . The resThe concentrations of Ni, Zn, and Cu were close to their background concentrations. It was therefore expected that there would not be significant pollution risks associated with Ni, Zn, and Cu. However, the pollution index of Pb, Cd, and Cr varied from the pre- to post-rainy seasons. The RI values showed that a large proportion of samples had moderate and high ecological risks associated with Cd. The high level of Cd risks agreed with the results of multiple studies in this region , 28, 31.The impact of land use types and crop rotations on the heavy metal accumulation has been investigated in many regions. Previous studies have shown high concentrations of heavy metals in cultivated land-use types rather than natural land-use types –10, 39. The precipitation was an important contributor to atmospheric transport at large spatial scales . The curAlthough we tried to address the potential limitations of our research, there were some issues that must be fully addressed in future studies. The sampling sites did not cover the topsoil in urban regions that may be more directly impacted by human-induced factors. The topsoil polluted by wastewater irrigation in some farmlands was also not considered in the current study. Recent studies have shown that high levels of heavy metal pollution resulted from wastewater irrigation and the application of wastewater sludge to farmland in some parts of this region , 40, 43.This study investigated the heavy metal concentrations in different land use types around the Beijing metropolis. The pollution levels and ecological risks associated with Cd were found to be higher than other heavy metals in all land use types. We also observed that concentrations of Pb, Ni, and Zn increased from the pre- to post-rainy season while those of Cd, Cr, and Cu decreased during this period. The results of this study indicated that the impact of land use on heavy metal risks was not statistically significant. We inferred that the atmospheric transport may play a key role in the distribution of heavy metals in this region. The further improvement of coupled models of atmospheric processes and industrial emission will be helpful to better understand the accumulation of heavy metals around the Beijing metropolis.S1 Tablehttps://figshare.com/articles/Supplementary_file_xlsx/3187756(XLSX)Click here for additional data file.