Additional file 1: of Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood

Figure S1. FACS staining of live/dead cells. A Representative images of FACS staining of live/dead cells of PB MNCs by four PB MNC isolation methods at day 0 or after 8 days. B Representative images of FACS staining of live/dead cells of PB MNCs at indicated time points. PB MNCs isolated with Ficoll method. (PPTX 99 kb

Additional file 2: of Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood

Figure S2. Differentiated PB iPSC clones did not express pluripotency markers OCT4, NANOG, TRA-1-60, and SSEA4. Representative images captured using Leica confocal microscope. (PPTX 292 kb

Romanian journal of psychoanalysis : = Revue Roumain de Psychanalyse

Course-dependent chemotherapy response of the T-ALL mice. When leukemic cells reached 1-5% in PB, mice received chemotherapy composed of CTX (100 mg/kg) and Ara-C (150 mg/kg) for a consecutive 1, 2, 3 and 4 days, respectively. (A) Median survival days post leukemic cell injection were 29, 39.5, 45 and 71.5 for the leukemia-only, one-day treated, two-day treated and three-day treated group, respectively (n = 6-10). For the four-day treated group, no mice died within the inspecting 90 days (n = 6). (B) Leukemic burden in PB of the four differently treating groups (n = 6). Data showed that longer the therapeutic course, longer the relapse-free period. The day on which leukemic cells showed up again in PB for the one-day, two-day and three-day treated groups were the 8th, 11th and 17th day post therapy, respectively. While for the four-day treated group, no appearance of relapse was detected within the inspecting 90 days. (C) White blood cell count in PB of the four differently treating groups (n = 6). Data showed that longer the therapeutic course, heavier the depression of white blood cells post therapy. (D) Platelet count in PB of the four differently treating groups (n = 6). Data showed that longer the therapeutic course, longer the suppression period of platelet in PB post therapy. All data were presented as meanÂąSEM. Statistical significance as: * p<0.05; ** p<0.01; *** p<0.001

Table2_High-expression of the innate-immune related gene UNC93B1 predicts inferior outcomes in acute myeloid leukemia.XLSX

Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy with dismal prognosis. Identification of better biomarkers remained a priority to improve established stratification and guide therapeutic decisions. Therefore, we extracted the RNA sequence data and clinical characteristics of AML from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression database (GTEx) to identify the key factors for prognosis. We found UNC93B1 was highly expressed in AML patients and significantly linked to poor clinical features (p < 0.05). We further validated the high expression of UNC93B1 in another independent AML cohort from GEO datasets (p < 0.001) and performed quantitative PCR of patient samples to confirm the overexpression of UNC93B1 in AML (p < 0.005). Moreover, we discovered high level of UNC93B1 was an independent prognostic factor for poorer outcome both in univariate analysis and multivariate regression (p < 0.001). Then we built a nomogram model based on UNC93B1 expression, age, FAB subtype and cytogenetic risk, the concordance index of which for predicting overall survival was 0.729 (p < 0.001). Time-dependent ROC analysis for predicting survival outcome at different time points by UNC93B1 showed the cumulative 2-year survival rate was 43.7%, and 5-year survival rate was 21.9%. The differentially expressed genes (DEGs) between two groups divided by UNC93B1 expression level were enriched in innate immune signaling and metabolic process pathway. Protein–protein interaction (PPI) network indicated four hub genes (S100A9, CCR1, MRC1 and CD1C) interacted with UNC93B1, three of which were also significantly linked to inferior outcome. Furthermore, we discovered high UNC93B1 tended to be infiltrated by innate immune cells, including Macrophages, Dendritic cells, Neutrophils, Eosinophils, and NK CD56dim cells. We also found UNC93B1 had a significantly positive correlation with CD14, CD68 and almost all Toll-like receptors. Finally, we revealed negatively correlated expression of UNC93B1 and BCL2 in AML and conjectured that high-UNC93B1 monocytic AML is more resistant to venetoclax. And we found high MCL-1 expression compensated for BCL-2 loss, thus, we proposed MCL-1 inhibitor might overcome the resistance of venetoclax in AML. Altogether, our findings demonstrated the utility of UNC93B1 as a powerful poor prognostic predictor and alternative therapeutic target.</p

Diterpenes from a Chinese Collection of the Brown Alga <i>Dictyota plectens</i>

Twenty-seven diterpenes of six chemical classes, including seven new diterpenes (<b>1</b>, <b>2</b>, <b>6</b>, <b>10</b>, <b>11</b>, <b>16</b>, and <b>19</b>), have been isolated from a collection of the brown alga <i>Dictyota plectens</i> from the South China Sea. The structures of the new diterpenes were elucidated by extensive spectroscopic analysis and by comparison with reported data. In the <i>in vitro</i> assays, <b>9</b>, <b>12</b>, <b>14</b>, <b>16</b>, and <b>22</b> showed inhibitory activity against HIV-1 replication with IC₅₀ values of 16.1–30.5 μM, compounds <b>5</b>, <b>13</b>, <b>24</b>, and <b>26</b> exhibited anti-H5N1 activity with inhibition rates of 50%–62% at 30.0 μM, and <b>12</b> and <b>24</b> also showed potent inhibition against LPS-induced NO production with inhibition rates of 90% and 86%, respectively, at 10.0 μM

CIB1 is an endogenous inhibitor of agonist-induced integrin aIIbb3 [alpha II b beta 3]activation

In response to agonist stimulation, the alpha IIb beta 3 integrin on platelets is converted to an active conformation that binds fibrinogen and mediates platelet aggregation. This process contributes to both normal hemostasis and thrombosis. Activation of alpha IIb beta 3 is believed to occur in part via engagement of the beta 3 cytoplasmic tail with talin; however, the role of the alpha IIb tail and its potential binding partners in regulating alpha IIb beta 3 activation is less clear. We report that calcium and integrin binding protein 1 (CIB1), which interacts directly with the alpha IIb tail, is an endogenous inhibitor of alpha IIb beta 3 activation; overexpression of CIB1 in megakaryocytes blocks agonist-induced alpha IIb beta 3 activation, whereas reduction of endogenous CIB1 via RNA interference enhances activation. CIB1 appears to inhibit integrin activation by competing with talin for binding to alpha IIb beta 3, thus providing a model for tightly controlled regulation of alpha IIb beta 3 activation

Similar ROS levels in BM cells between CL and WT mice.

<p>(<b>A</b>) CPR protein levels in bone marrow mononuclear cells (BMMNCs) and LKS⁺ of CL and WT mice were analyzed by Western blotting. (<b>B</b>) CPR mRNA expression in whole bone marrow cells (WBMCs), LKS⁺ and CD34⁻LKS⁺ were detected by qRT-PCR. Data shown are mean ± SEM (*P<0.05, **P<0.01, ***P<0.001, n3). (<b>C–D</b>) Representative FACS profiles (C) and quantification of the levels of ROS (D) in BM, LKS⁻ and LKS⁺ cells determined by DCF-DA mean fluorescence intensity with flow cytometry. Data shown are representative of two independent experiments.</p

Structurally Diverse Metabolites from the Soft Coral <i>Sinularia verruca</i> Collected in the South China Sea

Nineteen metabolites with diverse structures, including the rare pyrroloindoline alkaloid verrupyrroloindoline (<b>1</b>), the unprecedented highly fused benzosesquiterpenoid verrubenzospirolactone (<b>2</b>), the new asteriscane-type sesquiterpenoid 10-deoxocapillosanane D (<b>3</b>), and the two new cyclopentenone derivatives (4<i>S</i>*,5<i>S</i>*)-4-hydroxy-5-(hydroxymethyl)-2,3-dimethyl-4-pentylcyclopent-2-en-1-one (<b>4</b>) and (<i>S</i>)-4-hydroxy-5-methylene-2,3-dimethyl-4-pentylcyclopent-2-en-1-one (<b>5</b>), were isolated from a South China Sea collection of the soft coral <i>Sinularia verruca</i>. Eleven previously described marine metabolites (<b>7</b>–<b>15</b>, <b>18</b>, and <b>19</b>) were also obtained as well as three new EtOH-adduct artifacts (<b>6</b>, <b>16</b>, and <b>17</b>). The structures of the new compounds were elucidated by extensive spectroscopic analysis and by comparison with previously reported data. Compounds <b>4</b>, <b>5</b>, and <b>16</b> showed protection against the cytopathic effects of HIV-1 infection with EC₅₀ values of 5.8–34 μM, and <b>4</b>, <b>6</b>, and <b>16</b> exhibited inhibition against LPS-induced NO production with IC₅₀ values of 24–28 μM

No alteration of cell cycle in hematopoietic stem/progenitor cells of CL mice.

<p>(<b>A–C</b>) Cell cycle status of LKS⁺ and CD34⁻LKS⁺ from WT vs. CL mice. (<b>A</b>) Results are representatives of flow cytometry data in figures. (<b>B–C</b>) Data shown are quantification of mean cell cycle status ± SEM (n = 3–4) in LSK⁺ (B) and CD34⁻LKS⁺ (C).</p

Reconstitution upon third WBMCs transplantation.

<p>Third transplantation: 2×10⁵ CD45.2 cells sorted from 2^nd recipient WT or CL mice, plus 2×10⁵ CD45.1 WBMCs were transplanted into lethally irradiated (9.5 Gy) recipients (B6. SJL, CD45.1). Flow cytometry analysis of PB was performed after 8 weeks of transplantation. If the number of engrafted CD45.2⁺ cells in PB is greater than 1%, it was considered as positive reconstitution. Data were analyzed by χ², P<0.05.</p