Additional file 1: Table S1. of Iris ultrastructure in patients with synechiae as revealed by in vivo laser scanning confocal microscopy

Detailed structures observed in individual patients with synechaie. (DOCX 13 kb

pH-Responsive Artemisinin Derivatives and Lipid Nanoparticle Formulations Inhibit Growth of Breast Cancer Cells <em>In Vitro</em> and Induce Down-Regulation of HER Family Members

<div><p>Artemisinin (ART) dimers show potent anti-proliferative activities against breast cancer cells. To facilitate their clinical development, novel pH-responsive artemisinin dimers were synthesized for liposomal nanoparticle formulations. A new ART dimer was designed to become increasingly water-soluble as pH declines. The new artemisinin dimer piperazine derivatives (ADPs) remained tightly associated with liposomal nanoparticles (NPs) at neutral pH but were efficiently released at acidic pH's that are known to exist within solid tumors and organelles such as endosomes and lysosomes. ADPs incorporated into nanoparticles down regulated the anti-apoptotic protein, survivin, and cyclin D1 when incubated at low concentrations with breast cancer cell lines. We demonstrate for the first time, for any ART derivative, that ADP NPs can down regulate the oncogenic protein HER2, and its counterpart, HER3 in a HER2+ cell line. We also show that the wild type epidermal growth factor receptor (EGFR or HER1) declines in a triple negative breast cancer (TNBC) cell line in response to ADP NPs. The declines in these proteins are achieved at concentrations of NP109 at or below 1 µM. Furthermore, the new artemisinin derivatives showed improved cell-proliferation inhibition effects compared to known dimer derivatives.</p> </div

Effects of NP109 on the expression of selected proteins involved in cell proliferation, cell cycling, and apoptosis in BT474 (a–c) and MDA-MB-231 cells (d–f).

<p>Effects of NP109 on the expression of selected proteins involved in cell proliferation, cell cycling, and apoptosis in BT474 (a–c) and MDA-MB-231 cells (d–f).</p

Summary of IC₅₀ values calculated from MTT assays of ADPs and NPs on BT474 and MDA-MB-231 cells.

<p>Values represent average (±SD) calculated from three independent experiments. *Exceeded maximum concentration of assay.</p

Summary of loading and release efficiencies of the NPs.

<p>Values represent an average and standard deviation of three independent experiments read at λ = 263 nm.</p

Structures of artemisinin dimer succinate, ADPs 106–109 and ADPm109 the monomer analogue of compound ADP109.

<p>Structures of artemisinin dimer succinate, ADPs 106–109 and ADPm109 the monomer analogue of compound ADP109.</p

Requirement of Smad4 from Ocular Surface Ectoderm for Retinal Development

<div><p>Microphthalmia is characterized by abnormally small eyes and usually retinal dysplasia, accounting for up to 11% of the blindness in children. Right now there is no effective treatment for the disease, and the underlying mechanisms, especially how retinal dysplasia develops from microphthalmia and whether it depends on the signals from lens ectoderm are still unclear. Mutations in genes of the TGF-β superfamily have been noted in patients with microphthalmia. Using conditional knockout mice, here we address the question that whether ocular surface ectoderm-derived <i>Smad4</i> modulates retinal development. We found that loss of <i>Smad4</i> specifically on surface lens ectoderm leads to microphthalmia and dysplasia of retina. Retinal dysplasia in the knockout mice is caused by the delayed or failed differentiation and apoptosis of retinal cells. Microarray analyses revealed that members of Hedgehog and Wnt signaling pathways are affected in the knockout retinas, suggesting that ocular surface ectoderm-derived <i>Smad4</i> can regulate Hedgehog and Wnt signaling in the retina. Our studies suggest that defective of ocular surface ectoderm may affect retinal development.</p></div

Microarray analysis and Real-time qPCR showed the differentially expressed genes in control and <i>Smad4</i>-cKO mice.

<p><b>(A)</b> Overview of signaling pathway analysis defined by the SBS Analysis System (<a href="http://sas.ebioservice.com/" target="_blank">http://sas.ebioservice.com/</a>). <b>(B-C)</b> Charts showed the expression changes of genes in Hedgehog signaling pathway and Wnt signaling pathway detected by microarray within retina, respectively. Genes with more than 3 fold expression changes were used for further exploration. <b>(D)</b> Real-time qPCR was performed to detect the expression of Gli2, Gli3 and Wnt2b within retina at P0 and P8. The expression of Gli2, Gli3 and Wnt2b was down-regulated at P0 and P8. n = 4, *P<0.05. E, embryonic; P, postnatal; M, month.</p

Loss of <i>Smad4</i> affected the differentiation of retinal ganglion cells, bipolar cells, and Müller cells.

<p><b>(A-H)</b> Immunostaining was performed to label retinal ganglion cells (red) and bipolar cells (green) in P0, P9 and P14 <i>Smad4</i>-cKO and control mice. At P0, the number of ganglion cells in cKO retina was significantly more than that in control. At P9 and P14, the number of ganglion cells in cKO retina was significantly decreased. The bipolar cells showed delayed differentiation mainly at peripheral zone at P9. <b>(I-L)</b> Immunostaining was performed to label retinal ganglion cells (red) and Müller cells (green) in <i>Smad4</i>-cKO and control mice at P14 and 1M. In <i>Smad4</i>-cKO retina, the expression of GFAP was not observed from P9 through 1M after birth, indicating failed differentiation of Müller glia. E, embryonic; P, postnatal; M, month; GCL, ganglion cell layer; NBL, neuroblastic layer; INL, inner nuclear layer; ONL, outer nuclear layer; OLM, outer limiting membrane.</p

Loss of <i>Smad4</i> led to excessive proliferation of the retina cells at early development and abundant apoptosis later as assessed by pulse BrDU labeling and staining as well as TUNEL assay.

<p><b>(A-D)</b> Results of BrDU labeling followed by immunostaining were shown in <i>Smad4</i>-cKO mice and control mice at the nasal paracentral zone of P0 and P5. <b>(E-H)</b> TUNEL assay results were shown in <i>Smad4</i>-cKO mice and control mice at the nasal paracentral zone of P5 and P9. The <i>Smad4</i>-cKO retina exhibited more apoptosis at P5 and P9. <b>(I)</b> Quantification of total BrdU⁺ cells per retinal section in <i>Smad4</i>-cKO mice and control mice. From E14.5, the number of BrdU⁺ cells in the retina of cKO mice was significantly more than that of control. After birth, the number of BrdU⁺ retinal cells started to decrease in mice, with more decrease observed in cKO mice than in controls. n = 9, *P<0.05; **P<0.01. <b>(J)</b> Quantification of total apoptosis cells per retinal section in <i>Smad4</i>-cKO mice and control mice. In the first week after birth active retinal remodeling occurred and retinal cell apoptosis was detected in control mice. However, the <i>Smad4</i>-deficient retina exhibited more apoptosis than the control. n = 9. *P<0.05; **P<0.01. E, embryonic; P, postnatal; M, month; GCL, ganglion cell layer; NBL, neuroblastic layer; INL, inner nuclear layer; ONL, outer nuclear layer.</p