SETD7, H3K4me2, ZBTB20, and CDKN2D level in HCC was determined by TMA IHC.

<p>The two rows on the left indicate HE staining for HCC tumor tissues and paired ANLTs, whereas the two rows on the right indicate IHC staining for SETD7, H3K4me2, ZBTB20, and CDKN2D protein in HCC tumor tissues and paired ANLTs (<i>n</i> = 225). (Original magnified 100×; Inserted figures magnified 400×).</p

Increased Expression of <i>SETD7</i> Promotes Cell Proliferation by Regulating Cell Cycle and Indicates Poor Prognosis in Hepatocellular Carcinoma

<div><p>Purpose</p><p>To investigate the role of SET domain containing 7 (SETD7) in hepatocellular carcinoma (HCC) and determine whether SETD7 can be used as a predictor of overall survival in HCC patients.</p><p>Methods</p><p>mRNAs and proteins of <i>SETD7</i> and related genes in HCC tumor samples and paired adjacent non-tumorous liver tissues (ANLTs) (n = 20) or culture cells were determined by quantitative real-time PCR and Western blot. Cell proliferation and apoptosis with SETD7 knockdown SMMC-7721 cells or SETD7 overexpressed HepG2 cells were analyzed by CCK8 assay or flow cytometry. Gene expression alterations in SETD7 knockdown of SMMC-7721 cells were determined by digital gene expression (DGE) profiling. Defined data on patients (n = 225) with HCC were retrieved for the further study. Tissue microarrays (TMAs) were performed using paraffin tissues with tumor and ANLTs. SETD7 and related proteins were determined by TMAs immunohistochemistry. Statistical analyses were conducted to associate SETD7 expression with tumor features and patient outcomes, as well as related proteins expression.</p><p>Results</p><p>SETD7 expression was significantly higher in HCC tumor tissues than in ANLTs. SETD7 overexpression in vitro can promote HepG2 cell proliferation, whereas SETD7 knockdown can inhibit SMMC-7721 cell proliferation by regulating the cell cycle. SETD7 expression was significantly correlated with five genes expression. Increased SETD7 is associated with metastasis, recurrence, large tumor size, and poor tumor differentiation, and indicates poor prognosis in HCC patients.</p><p>Conclusions</p><p>SETD7 plays a critical role in HCC, and its immunohistochemistry signature provides potential clinical significance for personalized prediction of HCC prognosis.</p></div

<i>SETD7</i> expression changes liver cancer cell proliferation by regulating the cell cycle.

<p>(A) Western blot analysis was performed to detect the interference efficiency of siRNA after transfected with si-<i>SETD7</i>-1, si-<i>SETD7</i>-2, si-<i>SETD7</i>-3, or scramble RNA (NC) in SMMC-7721. (B) Western blot analysis was performed to detect <i>SETD7</i> expression in HepG2 transfected with pGV141-<i>SETD7</i> (OV-<i>SETD7</i>) or pGV141 plasmid (Control). (C, D) CCK8 array was used to assess proliferation in <i>SETD7</i> knockdown in SMMC-7721 cells or <i>SETD7</i> overexpression in HepG2 cells. *, P<0.05; **, P<0.01 by student’s t test. (E, F) Cell cycle was assessed in <i>SETD7</i> knockdown in SMMC-7721 cells or <i>SETD7</i> overexpression in HepG2 cells by flow cytometry assay.</p

Correlation between the SETD7 and clinicopatologic variables in HCC.

<p>Correlation between the SETD7 and clinicopatologic variables in HCC.</p

Kaplan–Meier survival analysis of the correlation between <i>SETD7</i> expression in patients with HCC and OS.

<p>Patients with high <i>SETD7</i> expression (solid line) exhibited decreased OS.</p

Correlation between the SETD7 and H3K4me2, ZBTB20, CDKN2D expression in HCC.

<p>Correlation between the SETD7 and H3K4me2, ZBTB20, CDKN2D expression in HCC.</p

Expression of <i>SETD7</i> in HCC tumor tissues, paired ANLTs and cell lines.

<p>(A) qRT-PCR analysis was performed to analyze <i>SETD7</i> expression in 20 pairs of HCC tumor tissues and ANLTs, the data shown are the mean of –ΔCT, and the expression of <i>SETD7</i> in HCC is significantly higher than that in ANLTs (P<0.05); (B) Western blot assay was performed to detect <i>SETD7</i> expression in 20 pairs of HCC tumor tissues and ANLTs; (C) Gray value of Western blot result (P<0.05); (D) Western blot assay was performed to analyze <i>SETD7</i> expression in normal hepatocyte cell line (HL-7702) and liver cancer cell lines (HepG2, SMMC-7721, QGY-7703, HCC-0010, and Bel-7404); (E). Typical IHC staining of SETD7 in HCC tumor tissues and ANLTs (with figures on the left magnified 100× and figures on the right magnified 400×); (F) Box plots indicate the IHC scores of SETD7 in HCC tumor tissues and ANLTs [mean, 3.204 (SD, 0.092) vs 1.427 (SD, 0.083), P<0.01].</p

Table1_The value of coordinated analysis of multimodal atherosclerotic plaque imaging in the assessment of cardiovascular and cerebrovascular events.docx

BackgroundAlthough atherosclerosis (AS) can affect multiple vascular beds, previous studies have focused on the analysis of single-site AS plaques.ObjectiveThe aim of this study is to explore the differences or similarities in the characteristics of atherosclerotic plaque found in the internal carotid artery, cerebral artery, and coronary artery between patients with atherosclerotic cardiovascular disease (ASCVD) and those without events.MethodsPatients aged ≥ 18 years who underwent both high-resolution vessel wall imaging (HR-VWI) and coronary computed tomography angiography (CCTA) were retrospectively collected and categorized into the ASCVD group and the non-event group. The plaques were then categorized into culprit plaques, non-culprit plaques, and non-event plaques. Plaque morphological data such as stenosis, stenosis grades, plaque length (PL), plaque volume (PV), minimal lumen area (MLA), enhancement grade, and plaque composition data such as calcified plaque volume (CPV), fibrotic plaque volume (FPV), fibro-lipid plaque volume (FLPV), lipid plaque volume (LPV), calcified plaque volume ratio (CPR), fibrotic plaque volume ratio (FPR), fibro-lipid plaque ratio (FLPR), lipid plaque volume ratio (LPR), intraplaque hemorrhage volume (IPHV), and intraplaque hemorrhage volume ratio (IPHR)were recorded and analyzed.ResultsA total of 44 patients (mean age 66 years, SD 9 years, 28 men) were included. In cervicocephalic plaques, the ASCVD group had more severe stenosis grades (p = 0.030) and demonstrated significant differences in LPV, LPR, and CPV (p = 0.044, 0.030, 0.020) compared with the non-event group. In coronary plaques, the ASCVD group had plaques with greater stenosis (p ConclusionThere is a consistent pattern of change in plaque characteristics between the cervicocephalic and coronary arteries in the same patient.</p

Puerarin Suppresses Proliferation of Endometriotic Stromal Cells Partly via the MAPK Signaling Pathway Induced by 17ß-estradiol-BSA

<div><h3>Background</h3><p>Puerarin is a major isoflavonoid compound extracted from <em>Radix puerariae</em>. It has a weak estrogenic action by binding to estrogen receptors (ERs). In our early clinical practice to treat endometriosis, a better therapeutic effect was achieved if the formula of traditional Chinese medicine included <em>Radix puerariae</em>. The genomic and non-genomic effects of puerarin were studied in our Lab. This study aims to investigate the ability of puerarin to bind competitively to ERs in human endometriotic stromal cells (ESCs), determine whether and how puerarin may influence phosphorylation of the non-genomic signaling pathway induced by 17ß-estradiol conjugated to BSA (E₂-BSA).</p> <h3>Methodology</h3><p>ESCs were successfully established. Binding of puerarin to ERs was assessed by a radioactive competitive binding assay in ESCs. Activation of the signaling pathway was screened by human phospho-kinase array, and was further confirmed by western blot. Cell proliferation was analyzed according to the protocol of CCK-8. The mRNA and protein levels of cyclin D1, Cox-2 and Cyp19 were determined by real-time PCR and western blotting. Inhibitor of MEK1/2 or ER antagonist was used to confirm the involved signal pathway.</p> <h3>Principal Findings</h3><p>Our data demonstrated that the total binding ability of puerarin to ERs on viable cells is around 1/3 that of 17ß-estradiol (E₂). E₂-BSA was able to trigger a rapid, non-genomic, membrane-mediated activation of ERK1/2 in ESCs and this phenomenon was associated with an increased proliferation of ESCs. Treating ESCs with puerarin abrogated the phosphorylation of ERK and significantly decreased cell proliferation, as well as related gene expression levels enhanced by E₂-BSA.</p> <h3>Conclusions/Significance</h3><p>Puerarin suppresses proliferation of ESCs induced by E₂-BSA partly via impeding a rapid, non-genomic, membrane-initiated ERK pathway, and down-regulation of Cyclin D1, Cox-2 and Cyp19 are involved in the process. Our data further show that puerarin may be a new candidate to treat endometriosis.</p> </div

<i>MiR-127</i> is induced from its promoter by DNA demethylation and histone deacetylase inhibition.

<p>(<b>A</b>) Schematic representation of the CpG island on the promoter region of the <i>miR-127</i> gene. (<b>B</b>) MiR-127 is highly induced by 5-Aza-CdR and PBA treatment. BRL-3A cells were treated with 5-Aza-CdR and/or PBA, and the miR-127 expression level was analyzed by qRT-PCR. U6 RNA expression was used as a loading control. 0.1A, 0.1 μmol/L 5-Aza-CdR; 1A, 1 μmol/L 5-Aza-CdR; 3A, 3 μmol/L 5-Aza-CdR; 0.1P, 0.1 mmol/L PBA; 1P, 1 mmol/L PBA; 3P, 3 mmol/L PBA; 0.1AP, combination of 0.1 μmol/L 5-Aza-CdR and 0.1 mmol/L PBA; 1AP, combination of 1 μmol/L 5-Aza-CdR and 1 mmol/L PBA; 3AP, combination of 3 μmol/L 5-Aza-CdR and 3 mmol/L PBA. Data from three independent experiments are shown as the means ± SD. (**<i>P</i><0.01). (<b>C</b>) The CpG island was hypermethylated 24 h after partial hepatectomy (PH). Genomic DNA was obtained from rat liver tissues 24 h after PH or sham operation (SH), the DNA methylation status was determined by bisulfite genomic sequencing (<i>P</i><0.01, Wilcoxon rank-sum test).</p