Changes in body weight, colon length and histological score of CD200^tg, WT CD200R1KO and CD200KO mice with DSS-induced chronic colitis.

<p>(A) Change in body weight. Percent weight loss from baseline (y-axis) is plotted versus time post initiation of first cycle of DSS (x-axis). Data points represent mean ± SD for 5 mice. (B) Change in colon length assessed on 8 days post initiation of the third cycle of DSS for same mice. Data are expressed as mean ± SD of five mice. (C) Change in histological score (again 8 days post start of the third cycle of DSS) for same mice. All independent experiments were performed three times, yielding similar results. * p < 0.05; ** p < 0.01; *** p < 0.001 compared with WT controls on the same day-see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.g001" target="_blank">Fig 1</a>.</p

Quantitation of staining of CD3⁺ and Foxp3⁺ cells in slides shown in Fig 5B and 5C.

<p>Quantitation of staining of CD3⁺ and Foxp3⁺ cells in slides shown in Fig <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.g005" target="_blank">5B and 5C</a>.</p

Effect of CD200 on expression of multiple cytokines and in DSS-induced acute colitis.

<p>Cytokines and chemokines were analyzed in colonic tissue from CD200^tg, WT, CD200R1KO and CD200KO mice with DSS-induced acute colitis. A and B, Real-time RT-PCR was performed using total RNA extracted from colonic tissue (5 samples/group) or LP cells (3 samples/group), with data normalized to the expression of GAPDH and HPRT in the same organ. Expression levels in CD200^tg, CD200R1KO and CD200KO mice (mean ± SD in 5 mice) are shown relative to that in WT at day 0 (designated as 1). * p < 0.05; ** p < 0.01; *** p < 0.001 compared to WT control (day 0). (C) Tissue explants from the four experimental groups were cultured and used for multi-analyte Elisarray. Again, relative expression levels of cytokine/chemokine (mean ± SD in 5 mice) in CD200^tg, CD200R1KO and CD200KO mice are shown compared to that in WT at day 0 (designated as 1), using the manufacturer’s software supplied. * p < 0.05; ** p < 0.01; *** p < 0.001, compared to this WT control (day 0). (D) Inflammatory cytokines measured by commercial ELISA (Biolegend: USA) in 36hr culture supernatants of colon-derived CD11c⁺, CD11b⁺, or CD4⁺ T cells (isolated by MACS columns) from WT, CD200^tg, CD200KO or CD200RKO mice 9 days post initiation of DSS treatment. Cells were obtained from a pool of 4 mice/group, and cultured without further stimulation for 36hrs in complete medium. In some cases, as shown, cells were cultured in the presence of CD200Fc (5mg/ml). Data show mean (<u>+</u>SD) of triplicate measurements. *, indicates p<0.05 compared with WT control; **indicates p<0.05 compared to equivalent group with no CD200Fc. E, Comparison of weight loss in 4 groups of DSS treated mice receiving anti-CD4mAb (right hand panel) or isotype control Ig (left hand panel)-see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.g001" target="_blank">Fig 1</a>. Data are for 4 mice/group, given 50mg Ig at days 4/7 post initiation of DSS treatment. Comparison of equivalent groups in right/left hand panels (Mann Whitney U-test) revealed no statistically significant differences.</p

Changes in body weight, colon length and histological score of CD200^tg, WT, CD200R1KO and CD200KO mice with DSS-induced acute colitis.

<p>(A) Change in body weight. Percent weight loss from baseline (y-axis) is plotted versus time post initiation of DSS treatment (x-axis). Data points represent mean ± SD for 5 mice. (B) Change in colon length assessed on day 9 following initiation of DSS treatment for the same same mice as in panel A. Data are expressed as mean ± SD of five mice. (C) Change in histological score (again day 9 post start of DSS) for mice of panel A. All independent experiments were performed three times, yielding similar results. * p < 0.05; ** p < 0.01; *** p < 0.001 compared with WT controls on the same day.</p

Primers used for real- time PCR.

<p>Primers used for real- time PCR.</p

Quantitation of staining of F4/80⁺ and GR1⁺ cells in slides shown in Fig 2B and 2C.

<p>Quantitation of staining of F4/80⁺ and GR1⁺ cells in slides shown in Fig <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.g002" target="_blank">2B and 2C</a>.</p

Effect of CD200 on histological changes and cell infiltration in colonic tissue from mice with DSS-induced acute colitis.

<p>Sections from colon (day 9 post start of DSS) of control (<i>a</i>), CD200^tg (<i>b</i>), WT (<i>c</i>), CD200R1KO (<i>d</i>) and CD200KO (<i>e</i>) mice were stained with (A) H&E, or with (B) rat anti-mouse F4/80, or (C) rat anti-mouse Gr1 mAbs, respectively. Percent staining in tissue is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.t001" target="_blank">Table 1</a>, counting 1000 cells for each group using 5 slides/group. (D) FACS staining with anti-F4/90 or anti-GR1 mAb using isolated LP cells obtained from the groups of mice documented above. Control cells were stained with Fluorochrome labeled mouse Ig.</p

Effect of CD200 on histological changes and cell infiltration in colons from mice with DSS-induced chronic colitis.

<p>Sections from colon (8 days post start of the third cycle of DSS) of control (<i>a</i>), CD200^tg (<i>b</i>), WT (<i>c</i>), CD200R1KO (<i>d</i>) and CD200KO (<i>e</i>) mice were stained with (A) H&E, or with (B) rat anti-mouse CD3, or (C) rat anti-mouse Foxp3 mAbs, respectively-see also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.g002" target="_blank">Fig 2</a>. Percent staining in tissue is shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146681#pone.0146681.t003" target="_blank">Table 3</a> using counts of 1000 cells for each group (5 slides/group).</p

The effect of CD200 on regulation of expression of CCR4 and Foxp3 in DSS-induced chronic colitis, and importance of CD25⁺Tregs and CD4⁺ T effector cells to DSS-induced weight loss.

<p>Foxp3, CCR4, CCL-17 and CCL-22 mRNA expression was analyzed in (A) colons and (B) LP cells by real-time RT-PCR from CD200^tg, WT, CD200R1KO and CD200KO mice with DSS-induced chronic colitis. All values were normalized to the expression of GAPDH and HPRT in the same organ with expression levels in CD200^tg, CD200R1KO and CD200KO mice expressed relative to that of WT (designated as 1). Data show mean ± SD for 5 mice/group. * p < 0.05; ** p < 0.01. (C) FACS staining for CD3⁺ and CD4⁺Foxp3⁺ (gated on CD3⁺ cells) in mesenteric lymph node (MLN) cells of mice in panels A and B. Data show typical staining patterns from pooled samples (3 mice/group). (D) Loss of protection from weight loss in DSS-induced chronic colitis in CD200^tg mice by treatment with anti-CD25mAb (Fig 7Da), and protection from weight loss in CD200RKO mice by treatment with anti-CD4 mAb (Fig 7Db). Data are shown for 4 mice/group (SD not shown to retain clarity in panels). Mice received 5mg antibody iv 1 day following completion of each DSS treatment. *p<0.05 (Mann-Whitney U-test) compared with equivalent groups not receiving mAb.</p

Datasheet1_Analysis of risk factors and construction of a prediction model for short stature in children.pdf

BackgroundShort stature in children is an important global health issue. This study aimed to analyze the risk factors associated with short stature and to construct a clinical prediction model and risk classification system for short stature.MethodsThis cross-sectional study included 12,504 children aged 6–14 years of age from 13 primary and secondary schools in Pingshan District, Shenzhen. A physical examination was performed to measure the height and weight of the children. Questionnaires were used to obtain information about children and their parents, including sex, age, family environment, social environment, maternal conditions during pregnancy, birth and feeding, and lifestyle. The age confounding variable was adjusted through a 1 : 1 propensity score matching (PSM) analysis and 1,076 children were selected for risk factor analysis.ResultsThe prevalence of short stature in children aged 6–14 years was 4.3% in the Pingshan District, Shenzhen. The multivariate logistic regression model showed that the influencing factors for short stature were father's height, mother's height, annual family income, father's level of education and parents’ concern for their children's height in the future (P ConclusionThis study analyzed the risk factors for short stature in children and constructed a nomogram prediction model and a risk classification system based on these risk factors, as well as providing short stature screening and assessment individually.</p