Table_2_The cross-sectional relationship between vitamin C and high-sensitivity C-reactive protein levels: insights from NHANES database.DOCX

BackgroundAscorbic acid or vitamin C has antioxidant and anti-inflammatory properties that may impact markers of inflammation like C-reactive protein (CRP). However, studies specifically on vitamin C and high-sensitivity CRP (hs-CRP) have been scarce.MethodsWe conducted a cross-sectional analysis of the National Health and Nutrition Examination Survey 2017–2018 dataset including 5,380 U.S. adults aged ≥20 years. Multiple regression models examined the relationship between plasma vitamin C and serum hs-CRP while adjusting for potential confounders. Stratified analyses and curve fitting assessed effect modification and nonlinearity.ResultsAn inverse association was found between plasma vitamin C and serum hs-CRP overall (β = −0.025, 95% CI: −0.033 to −0.017, p ConclusionThe results showed a non-linear negative correlation between vitamin C levels and hs-CRP in adults. These results suggest vitamin C intake may reduce inflammation and cardiovascular risk, but only up to 53.1 μmol/L plasma vitamin C.</p

Table_1_The cross-sectional relationship between vitamin C and high-sensitivity C-reactive protein levels: insights from NHANES database.docx

BackgroundAscorbic acid or vitamin C has antioxidant and anti-inflammatory properties that may impact markers of inflammation like C-reactive protein (CRP). However, studies specifically on vitamin C and high-sensitivity CRP (hs-CRP) have been scarce.MethodsWe conducted a cross-sectional analysis of the National Health and Nutrition Examination Survey 2017–2018 dataset including 5,380 U.S. adults aged ≥20 years. Multiple regression models examined the relationship between plasma vitamin C and serum hs-CRP while adjusting for potential confounders. Stratified analyses and curve fitting assessed effect modification and nonlinearity.ResultsAn inverse association was found between plasma vitamin C and serum hs-CRP overall (β = −0.025, 95% CI: −0.033 to −0.017, p ConclusionThe results showed a non-linear negative correlation between vitamin C levels and hs-CRP in adults. These results suggest vitamin C intake may reduce inflammation and cardiovascular risk, but only up to 53.1 μmol/L plasma vitamin C.</p

Candidate genes involved in BMP signaling in <i>Drosophila</i> S2 cells.

<p>*Rank shows the order of loss-of-BMP signaling due to RNAi.</p><p>Candidate genes involved in BMP signaling in <i>Drosophila</i> S2 cells.</p

Role of TRPM7 Channels in Hyperglycemia-Mediated Injury of Vascular Endothelial Cells

<div><p>This study investigated the change of transient receptor potential melastatin 7 (TRPM7) expression by high glucose and its role in hyperglycemia induced injury of vascular endothelial cells. Human umbilical vein endothelial cells (HUVECs) were incubated in the presence or absence of high concentrations of D-glucose (HG) for 72h. RT-PCR, Real-time PCR, Western blotting, Immunofluorescence staining and whole-cell patch-clamp recordings showed that TRPM7 mRNA, TRPM7 protein expression and TRPM7-like currents were increased in HUVECs following exposure to HG. In contrast to D-glucose, exposure of HUVECs to high concentrations of L-glucose had no effect. HG increased reactive oxygen species (ROS) generation, cytotoxicity and decreased endothelial nitric oxide synthase protein expression, which could be attenuated by knockdown of TRPM7 with TRPM7 siRNA. The protective effect of silencing TRPM7 against HG induced endothelial injury was abolished by U0126, an inhibitor of the extracellular signal-regulated kinase signaling pathway. These observations suggest that TRPM7 channels play an important role in hyperglycemia-induced injury of vascular endothelial cells.</p> </div

Prognostic impact of PRDM16 expression in acute myeloid leukemia with normal cytogenetics

Cytogenetically normal acute myeloid leukemia (CN-AML) is a heterogeneous disease with variable clinical outcomes. The identification of potential biomarkers to better classify the patients with unfavorable prognoses who may require more aggressive therapies is an emergent demand. PRDM16 is a transcriptional cofactor and histone methyltransferase, playing a critical role in maintaining hematopoietic stem cells, and MLL fusion-induced leukemogenesis. However, the prognostic value of PRDM16 in CN-AML is still unclear. We retrospectively analyzed the PRDM16 expression and its association with gene mutations in CN-AML. Then the prognostic value of PRDM16 and its comparison with WT1 were analyzed. The results showed that about 73.6% of CN-AML patients harbored higher expression of PRDM16 than the healthy controls. Furthermore, CN-AML patients with high PRDM16 expression had a lower survival rate than the low PRDM16 expression group (50.5% vs. 83.3%, p = 0.0339). Interestingly, hemopoietic stem cell transplantation significantly improved the prognosis of CN-AML with high PRDM16 expression but not those with low PRDM16 expression. In terms of molecular genetics, high PRDM16 expression was significantly associated with a lower rate of CEBPA mutation (p = 0.01) and a higher rate of FLT3-ITD and DNMT3A mutation (p = 0.032 and p = 0.004, respectively). In addition, PRDM16 expression was significantly correlated with WT1 expression in CN-AML (r = 0.7, p  PRDM16 is significantly associated with FLT3-ITD and DNMT3A mutation and WT1 expression and serves as a potential prognostic biomarker in CN-AML.</p

Ter94 is involved in BMP signaling in <i>Drosophila</i> S2 cells.

<p>(A) BMP signals are regulated by Ter94 in S2 cells. The loss of BMP signaling caused by <i>Ter94</i> (3′-UTR) RNAi was restored by the ectopic expression of <i>Ter94</i>. <i>pAW-Ter94</i> was used for the expression of <i>Ter94</i> in S2 cells; whereas <i>pBS</i> served as a control vector. Tubulin was used for a loading control. (B) The relative intensities of pMad signals (compared to the pMad signal of ds-<i>lacI</i>) were measured with Western blotting. pMad signals were normalized by Tubulin expression; ds-<i>lacI</i> intensity was set at 100%. ** indicates P<0.01. (C) The relative protein level of Ter94 (compared to the protein level of ds-<i>lacI-</i>treated cells) was quantified. The Ter94 level was normalized by Tublin expression; ds-<i>lacI</i> intensity was set at 100%. ** indicates P<0.01. (D) Ter94 is involved in BMP signaling upstream of Smurf or independent of Smurf in S2 cells. Tubulin was used for a loading control. (E) The relative intensities of pMad signaling (compared to the pMad signal of ds-<i>lacI</i>) were measured with Western blotting. pMad signals were normalized by Tubulin expression; ds-<i>lacI</i> intensity was set at 100%. * indicates P<0.05 and ** indicates P<0.01. Note that the pMad signal of the double knockdown of <i>Ter94</i> and <i>Smurf</i> is similar to that of <i>Ter94</i> RNAi. (F) The loss of BMP signaling caused by <i>Ter94</i> RNAi was restored by the ectopic expression of <i>hVCP</i>. The relative intensities of pMad signaling (compared to the pMad signal of ds-<i>lacI</i>) were measured with Western blotting. <i>pAW-Ter94</i> or <i>pAW-hVCP</i> was used for the expression of <i>Ter94</i> or <i>hVCP</i> in S2 cells, respectively. pMad signals were normalized by <i>Flag-Mad</i> expression; ds-<i>lacI</i> intensity was set at 100%. ** indicates P<0.01. dsRNA against <i>lacI</i> served as a control to rule out any specific effects caused by RNAi transfection, and <i>put</i> was used to validate that its knockdown inhibits pMad signals.</p

VCP is involved in BMP signaling in mammalian cells.

<p>(A) Alkaline phosphatase (ALP) assay in C2C12 cells. BMP6-induced ALP activity was abolished by the knockdown of Smad1 or VCP. ** indicates P<0.01. (B) BMP-induced activity of the luciferase reporter (BRE-Luc) in KS483 cells. BMP6-induced luciferase activity was inhibited by the overexpression of VCP^R155H but was activated by the overexpression of VCP^WT. * indicates P<0.05.</p

Effect of HG on TRPM7 protein expression in HUVECs.

<p>(A) Representative immunoblots showing TRPM7 protein expression in HUVECs with or without HG (30 mM) for 72h. (B) The corresponding bar graphs showing relative expression of TRPM7 protein normalized to beta-actin. (C) Representative TRPM7-like currents recorded in HUVECs cultured in control or HG (30 mM) for 72h. (D) TRPM7-like current density. **<i>p</i><0.01 vs. HG; *<i>p</i><0.05 vs. control. n=4 for immunoblotting and 17-18 cells patched for current recording. </p

Ter94 is involved in BMP signaling <i>in vivo</i>.

<p>(A) <i>Ter94</i> alleles and <i>dpp</i> show genetic interactions. The <i>dpp</i> null mutant is haploinsufficient. In a <i>Smurf</i> mutant background, the <i>dpp</i> heterozygote is partially viable. In <i>Ter94</i> and <i>Smurf</i> double mutants, the viability of the <i>dpp</i> heterozygote is less than that of the <i>Smurf</i> mutant alone. (B–H) A dorsal view of <i>race</i> mRNA in early stage-6 embryos. <i>dpp</i> null (B), <i>dpp</i> heterozygote (C), wild-type phenotype (D), three copies of Dpp (+/P[<i>dpp</i>-P23]: E), transheterozygous for <i>dpp</i> null and <i>Smurf</i> mutant alleles (<i>dpp^H46 Smurf⁺/dpp⁺ Smurf^C15</i>: F), transheteorzygous for <i>dpp</i> null, <i>Smurf</i> mutant and <i>Ter94</i> mutant alleles (<i>dpp^H46 Ter94⁺ Smurf⁺/dpp⁺ Ter94^EY03468 Smurf^C15</i>: G, <i>dpp^H46 Ter94⁺ Smurf⁺/dpp⁺ Ter94^k15002 Smurf^C15</i>: H). (I) Number of <i>race</i> staining cells in early stage-6 embryos. ** indicates <i>P</i><0.01 (compared with wild-type phenotype <i>yw</i>).</p

Imaging-based assay of BMP signaling for RNAi screening in <i>Drosophila</i> S2 cells.

<p>(A) Imaging-based analysis of BMP signals in <i>Drosophila</i> S2 cells. Dpp, Scw and Flag-tagged Mad were co-transfected in S2 cells together with dsRNA in 384-well plates. Mad expressions were detected with anti-Flag/Alexa 488 anti-mouse IgG antibodies, and BMP signals were detected with anti-pMad/Alexa 647 anti-rabbit IgG antibodies. dsRNA against <i>gfp</i> served as a control to rule out any nonspecific effects caused by RNAi transfection, and the BMP type II receptor <i>put</i> served to validate that its knockdown inhibits pMad expression. (B) Imaging analysis of the BMP signals in S2 cells reflects the relative intensities that were regulated by various RNAi approaches (ds-<i>gfp</i>, ds-<i>sax</i>, ds-<i>tkv</i>, and ds-<i>put</i>). The relative intensities of BMP signals by RNAi from four independent experiments were measured with imaging-based analysis. The intensity of ds-<i>gfp</i> was set at 100%. ** indicates P<0.01 (ds-<i>sax</i>, ds-<i>tkv</i>, or ds-<i>put</i> was compared to ds-<i>gfp</i>). (C) The protocol for genome-wide RNAi screening of BMP signaling. The Arrayscan program enabled to calculate the number of DAPI (channel 1) as a total cell number; the number of Flag-Mad-positive cells (channel 2) within the DAPI-positive cells was counted as the Mad-positive cells, and then the number of pMad-positive cells (channel 3) within the Flag-positive cells was counted as the pMad-positive cells. The relative intensity of each sample was calculated as a percentage of pMad staining against Mad (Flag)-positive cells.</p