Electrospun Polycaprolactone/Polyvinylidene Fluoride Composite Nanofibers for Fabricating Artificial Conduits for Ureteral Stricture Treatment

The ureter is a tube that carries urine excreted from the kidneys from the renal pelvis to the bladder and is of small diameter at (2–10) mm. Ureteral stricture refers to a condition in which the ureter is obstructed. Similar to arteriosclerosis and esophageal cancer patients, ureteral stenosis can also be treated by re-expanding the stenotic area using a stent or using a ureteral splint. However, even after treatment, the recurrence rate of ureteral stenosis is very high, over 80%, and has an average replacement cycle of (3–6) months. During the initial stent placement and replacement process, the patient feels great pain and, during the stent placement, also feels uncomfortable. In this study, multilayer membrane fabrication was easily performed by installing several polymer solutions at once using a high-speed electrospinning device that simultaneously operates four nozzles, not the electrospinning system commonly used in the past. In addition, we show that by using various sizes of collectors, it is possible to manufacture a small-diameter catheter of (3–10) mm similar to the actual ureter size. The functions of the electrospun PCL(CEO)/PVDF multilayer membrane manufactured in this study were confirmed according to the additives added to each layer and the polymer used. It was confirmed that the CEO added first had an antibacterial effect. In addition, it was confirmed that piezoelectric response was detected on both sides when stimulation was applied to the electrospun PCL(CEO)/PVDF multilayer membrane by the PVDF layer. The artificial catheter manufactured in this study was added with cinnamon essential oil, a representative antibacterial substance, to PCL, a biocompatible polymer, to prevent the accumulation of foreign substances and the formation of stones due to the formation of bio-sludge, which is the main cause of ureteral stricture. In addition, PVDF, a piezoelectric polymer, is coated on the PCL layer so that the PVDF layer is in contact with the actual ureter muscle. When stimulated by the flow of bodily fluid or muscle movement, PVDF piezoelectricity generates an electric current, which can have the effect of stimulating and treating malignant tumors in the ureter. The artificial conduit manufactured in this study is expected to apply both to the ureter and to blood conduits and various tissues in the body

Ru(II)-Catalyzed Site-Selective Hydroxylation of Flavone and Chromone Derivatives: The Importance of the 5‑Hydroxyl Motif for the Inhibition of Aurora Kinases

An efficient protocol for Ru­(II)-catalyzed direct C–H oxygenation of a broad range of flavone and chromone substrates was developed. This convenient and powerful synthetic tool allows for the rapid installation of the hydroxyl group into the flavone, chromone, and other related scaffolds and opens the way for analog synthesis of highly potent Aurora kinase inhibitors. The molecular docking simulations indicate that the formation of bidentate H-bonding patterns in the hinge regions between the 5-hydroxyflavonoids and Ala213 was the significant binding force, which is consistent with experimental and computational findings

GS-ECFC-mediated neovascularization in border zone of LV infarct at 28 days post-MI.

<p>(A) IF staining for CD 31 (green) in ischemic heart tissue at 28 days after ECFCs were grafted as genistein stimulate-ECFC (GS-ECFC) or CTRL (control untreated ECFC). (Scale bar: 50 µm). (B) The bar graph shows the quantification of CD31⁺ capillary density. (C) Engraftment of ECFC (HNA+, red fluorescence) into vascular structures (CD31 staining for capillaries green fluorescence) is seen as yellow structures. <b>Insert</b> is higher magnification of the <b>yellow-boxed area</b>. (D) The bar graph shows the quantification of HNA⁺ cells associated with CD31+ vasculature. (E) Engraftment of ECFC (HNA+, green fluorescence) into arteriole structures (α-SMA staining for arterioles red fluorescence) is seen as yellow structures. <b>Insert</b> is higher magnification of the <b>yellow-boxed area</b>. (F) The bar graph shows the quantification of HNA⁺ cells associated with α-SMA + arterioles. HPF indicates high-power field. (n = 6) *P<0.05 vs. CTRL (indicates control genistein untreated ECFC).</p

Genistein Promotes Endothelial Colony-Forming Cell (ECFC) Bioactivities and Cardiac Regeneration in Myocardial Infarction

<div><p>Although stem cell-mediated treatment of ischemic diseases offers significant therapeutic promise, the limitation in the therapeutic efficacy of transplanted stem cells <i>in vivo</i> because of poor engraftment remains a challenge. Several strategies aimed at improving survival and engraftment of stem cells in the ischemic myocardium have been developed, such as cell transplantation in combination with growth factor delivery, genetic modification of stem cells, and/or cell therapy using scaffolds. To improve therapeutic efficacy, we investigated the effects of genistein on the engraftment of transplanted ECFCs in an acute myocardial ischemia model. <i>Results</i>: We found that genistein treatment enhanced ECFCs' migration and proliferation, which was accompanied by increases in the expression of ILK, α-parvin, F-actin, and phospholylation of ERK 1/2 signaling. Transplantation of genistein-stimulates ECFCs (GS-ECFCs) into myocardial ischemic sites <i>in vivo</i> induced cellular proliferation and secretion of angiogenic cytokines at the ischemic sites and thereby enhanced neovascularization and decreased myocardial fibrosis as well as improved cardiac function, as shown by echocardiography. Taken together, these data suggest that pretreatment of ECFCs with genistein prior to transplantation can improve the regenerative potential in ischemic tissues, providing a novel strategy in adult stem cell therapy for ischemic diseases.</p></div

ERK1/2-mediated genistein-induced ECFC proliferation and survival in the border zone of the left ventricular (LV) infarct at 3 days after myocardial infarction (MI).

<p>(A) ECFCs were pretreated with U0126 (ERK1/2 inhibitor, 10⁻⁶ M) for 30 min prior to 12 h of genistein treatment and then washed with PBS, fixed, stained, and analyzed by flow cytometry. Gates were manually configured to determine the percentage of cells in S phase based on DNA content (n = 5). *P<0.05 vs. CTRL (indicates control genistein untreated ECFC), **P<0.05 vs. genistein stimulate-ECFC (GS-ECFC). ECFCs were pretreated with U0126 for 30 min prior to a 12 h genistein (10⁻¹⁰ M) treatment, and the cells were transplanted into the ischemic region. (B) Proliferating cell nuclear antigen (PCNA) staining to detect ECFC proliferation. PCNA+ cell zone, <b>yellow-boxed area.</b> (Scale bar: 100 µm). (C) Quantification of PCNA-positive cells at 3 d after MI. (D) Co-immunofluorescent staining to detect proliferation (Ki67 [proliferation marker, red] and of hECFCs [human nuclear antigen (HNA)-positive cells, green] and DAPI [blue] for nuclear staining). <b>Arrows</b> indicate Ki67+ HNA+ DAPI+ cells. (E) Quantitative analysis of Ki67/HNA/DAPI triple-positive cells at 3 days after MI. (F) Coimmunofluorescent staining to detect apoptosis (caspase-3, apoptosis marker, green) and of hEPCs (HNA-positive cells, red) and DAPI (blue) by nuclear staining. <b>Arrows</b> indicate caspase-3+ HNA+ DAPI+ cells. (Scale bar: 20 µm). (G) Quantitative analysis of caspase-3/HNA/DAPI triple-positive cells at 3 days after MI. (n = 6) *P<0.05 vs. CTRL (indicates control genistein untreated ECFC), **P<0.05 vs. genistein stimulate-ECFC (GS-ECFC).</p

Schematic representation of the mechanism underlying the <i>P</i>. <i>lactiflora</i> induced LIF expression in the endometrium and its relevance.

<p><i>P</i>. <i>lactiflora</i> induces LIF expression through the activation of p38 and MEK/ERK pathways, leading to an increase in the trophoblast adhesion to the endometrium. Inhibition of LIF expression blocks the expression of adhesion molecules, such as integrin β3 and β5, and adhesion of the trophoblast to the endometrium.</p

ILK, α-parvin and F-actin mediated genistein-induced ECFC migration.

<p>(A) ECFCs were treated with genistein for 0–24 h, and ILK, α-parvin and F-actin was detected by western blotting. (B) ECFCs were transfected with ILK, α-parvin, and TRIOBP (F-actin) small interfering RNA (siRNA) (ILK, α-parvin, and TRIOBP-specific siRNA; 200 pmol) for 24 h before genistein treatment and staining with Calcein AM. Fluorescence in the analytical zone was quantified with a plate reader. *P<0.05 vs. CTRL (indicates control genistein untreated ECFCs), **P<0.05 vs. genistein. (C) ECFCs were transfected with ILK siRNA (ILK-specific siRNA; 200 pmol) for 24 h before genistein (10⁻¹⁰ M) treatment, and the cells were injected into the tail veins of mice 30 min after left anterior descending (LAD) artery ligation. Staining of ECs with isolectin B4 (green) showed human nuclei antibody (HNA) (red)-positive cell incorporation into the border zone of left ventricular (LV) infarct at 3 days after myocardial infarction (MI) (Scale bar: 20 µm). <b>Inset</b> in higher magnification of the <b>yellow-boxed area</b>. <b>Arrows</b> indicate of isolectin B4+HNA+DAPI+cells. (D) The bar graph shows quantitative analysis of the number of HNA+cells associated with isolectin B4+vasculature (n = 5). HPF indicates high-power field. *P<0.05 vs. CTRL (indicates control genistein untreated ECFC), **P<0.05 vs. genistein stimulate-ECFC (GS-ECFC).</p

Effect of genistein on ECFC migration and proliferation.

<p>(A) ECFCs were incubated for 12 h with various concentrations of genistein (10⁻¹⁰–10⁻⁵ M) and then stained with 5 µM Calcein AM. Fluorescence was quantified with a plate reader. (B) The <i>in vitro</i> ECFC wound-healing motility assay was performed in the absence and presence of genistein. Ten fields per plate were examined. (Scale bar: 100 µm). (C) ECFCs were incubated for 12 h with various concentrations of genistein, and CDK 2, cyclin E, CDK 4, and cyclin D₁ were assessed by western blotting. (D) ECFCs were incubated for 12 h with various concentrations of genistein, and assessed by MTT. (E) ECFCs were treated with genistein and then washed with PBS, fixed, stained, and analyzed by flow cytometry. Gates were manually configured to determine the percentage of cells in S phase based on the DNA content (<i>n</i> = 5). *<i>P</i><0.05 vs. CTRL (indicates control genistein untreated ECFCs).</p

<i>Paeonia lactiflora</i> Enhances the Adhesion of Trophoblast to the Endometrium via Induction of Leukemia Inhibitory Factor Expression

<div><p>In the present study, we investigated the role of <i>Paeonia lactiflora Pall</i>. extract on embryo implantation <i>in vitro</i> and <i>in vivo</i>. A polysaccharides depleted-water extract of <i>P</i>. <i>lactiflora</i> (PL-PP) increased LIF expression in human endometrial Ishikawa cells at non-cytotoxic doses. PL-PP significantly increased the adhesion of the human trophectoderm-derived JAr spheroids to endometrial Ishikawa cells. PL-PP-induced LIF expression was decreased in the presence of a p38 kinase inhibitor SB203580 and an MEK/ERK inhibitor U0126. Furthermore, endometrial LIF knockdown by shRNA reduced the expression of integrins β3 and β5 and adhesion of JAr spheroids to Ishikawa cells. <i>In vivo</i> administration of PL-PP restored the implantation of mouse blastocysts in a mifepristone-induced implantation failure mice model. Our results demonstrate that PL-PP increases LIF expression via the p38 and MEK/ERK pathways and favors trophoblast adhesion to endometrial cells.</p></div

Expression of adhesion molecules in PL-PP treated Ishikawa cells.

<p><b>(A)</b> Ishikawa cells were treated with or without PL-PP (50 μg/mL) for 24 h, and total RNA was extracted. The expression levels of <i>LIF</i>, <i>ITGAV</i>, <i>ITGB1</i>, <i>ITGB3</i>, <i>ITGB4</i>, <i>ITGB5</i>, <i>ICAM-1</i>, <i>L-selectin</i>, <i>E-cadherin</i>, and <i>CD44</i> mRNA were analysed by RT-PCR. β-actin was used as an internal control. <b>(B)</b> The pLKO.1 –or shLIF—transfected Ishikawa cells were treated with PL-PP (50 μg/mL) for 24 h. Total RNA was extracted and <i>ITGB3</i> and <i>ITGB5</i> mRNA expression levels were measured by RT-PCR. β-actin was used as an internal control.</p