Groups and treatments.

a<p>: Administrated once daily by oral gavage from 1^st–20^th day;</p>b<p>: Administrated intraperitoneal 30 min before a training course of Y-maze test at the 20^th day;</p>c<p>: body weight.</p><p>Groups and treatments.</p

Effects of combined proanthocyanidins extracted from lotus seedpod (LSPC) and <i>Lactobacillus casei-01</i> (LC) on scopolamine-induced memory impairment in Y-maze test.

<p>Groups without any same letters above the bars signify statistically significant differences (P<0.05). CON, SCOP, Piracetam and LC is control, vehicle scopolamine control, positive drug control and <i>Lactobacillus casei-01</i> group, respectively. L-LSPC and H-LSPC is low and high dose of LSPC group, respectively. L-LSPC+LC is L-LSPC and LC combination group. H-LSPC+LC is H-LSPC and LC combination group.</p

Effects of combined proanthocyanidins extracted from lotus seedpod (LSPC) and <i>Lactobacillus casei-01</i> (LC) on brain malondialdehyde (MDA) and total antioxidant capacity (TAOC) levels, total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), myeloperoxidase (MPO) and acetylcholinesterase (AchE) activities, ratio of T-SOD to MDA (T-SOD/MDA) and ratio of GSH-Px to T-SOD (GSH-Px/T-SOD) in scopolamine-induced amnesia mice.

<p>Notes: Means in the same column with different superscript are significantly different (P<0.05), while sharing any same letters signify insignificant differences. CON, SCOP, Piracetam and LC is control, vehicle scopolamine control, positive drug control and <i>Lactobacillus casei-01</i> group, respectively. L-LSPC and H-LSPC is low and high dose of LSPC group, respectively. L-LSPC+LC is L-LSPC and LC combination group. H-LSPC+LC is H-LSPC and LC combination group.</p><p>Effects of combined proanthocyanidins extracted from lotus seedpod (LSPC) and <i>Lactobacillus casei-01</i> (LC) on brain malondialdehyde (MDA) and total antioxidant capacity (TAOC) levels, total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), myeloperoxidase (MPO) and acetylcholinesterase (AchE) activities, ratio of T-SOD to MDA (T-SOD/MDA) and ratio of GSH-Px to T-SOD (GSH-Px/T-SOD) in scopolamine-induced amnesia mice.</p

Effects of combined proanthocyanidins extracted from lotus seedpod (LSPC) and <i>Lactobacillus casei-01</i> (LC) on the mRNA level of neural nitric oxide synthase (nNOS).

<p>Groups without any same letters above the bars signify statistically significant differences (P<0.05). CON, SCOP, Piracetam and LC is control, vehicle scopolamine control, positive drug control and <i>Lactobacillus casei-01</i> group, respectively. L-LSPC and H-LSPC is low and high dose of LSPC group, respectively. L-LSPC+LC is L-LSPC and LC combination group. H-LSPC+LC is H-LSPC and LC combination group.</p

Additional file 1 of A novel AML1-ETO/FTO positive feedback loop promotes leukemogenesis and Ara-C resistance via stabilizing IGFBP2 in t(8;21) acute myeloid leukemia

Additional file 1: Fig. S1. AML1-ETO promotes expression of FTO via PU.1. (A) Comparison of overall survival of patients with de novo t(8;21) AML (n = 26) using the Kaplan–Meier method grouped by the expression of FTO (high vs. low). p value was evaluated using the log-rank test. (B) Kaplan–Meier analysis of event-free survival (left) and overall survival (right) of patients with AML (n = 344, data from GSE6891) based on the expression of FTO. (C) Comparison of the expression of FTO in Kasumi-1 cells with or without AML1-ETO knockdown (shAE vs. shNS) detected by RNA-seq in the GSE115121 data set. (D) ChIP-seq of GSE65427 depicting FTO loci in Kasumi-1 cells targeting C- terminus of ETO (upper panel) and N-terminus of AML1(lower panel), representing AML1-ETO peaks on FTO. (E) Schematic diagrams showing the amplified regions on the promoter of SPI1 for the ChIP-qPCR showed in Fig. 1I and Fig. S1F. The location of targeted amplified region (named ‘Target’) and negative control site (NC) are indicated with blue horizontal lines. The red triangle indicates the location of peak summit of AML1-ETO on the promoter of SPI1 detected by the ChIP-seq of GSE65427. (F) ChIP-qPCR assays showing no direct binding of AML1 or ETO within 200 bp upstream of the SPI1 promoter in SKNO-1-siAE cells. (G and H) Pearson correlation of the expression of FTO and SPI1 in (G) normal blood tissues from the Genotype-Tissue Expression Project (GTEx, n = 444) or (H) BM samples of patients with AML from TCGA database (n = 173). (I) Sequences of the 4 FTO promoter fragments (the P1 to P4 showed in Fig. 1P) and putative PU.1 binging sites (sites 1, 2, and 3). Fig. S2. FTO upregulated AML1-ETO in a m6A-dependent manner. (A–C) The level of AML1-ETO mRNA detected by qPCR in SKNO-1 and Kasumi-1 cells (A) transduced with wild-type FTO (wt-FTO), mutant FTO (mut-FTO), or mock vectors; (B) transduced with FTO-knockdown (shFTO#1 and shFTO#2) or scramble shRNA (shNS) vectors; (C) treated with DMSO or FB23-2 treatment for 72 h. Fig. S3. Oncogenic role of FTO in t(8;21) AML cells and AML1-ETO9a driven AML mice. (A) Effects of forced expression or knockdown of FTO on cell cycle in SKNO-1 and Kasumi-1 cells. (B) Effects of forced expression or knockdown of FTO on colony-forming capacity of Kasumi-1 cells. (C) The effect of FTO knockdown on differentiation of Kasumi-1 cells. The percentage of CD11b+ cells was quantified (right panel). (D) Wright-Giemsa staining of Kasumi-1 cells with or without FTO knockdown. (E) Spleen size in AML1-ETO9a-driven AML mice with or without Fto knockdown or treatment with DMSO or FB23-2 (6 mg/kg) 7 weeks after transplantation (n = 6 for each group). (F) Spleen weight of AML1-ETO9a-driven AML mice from (E). (G) Wright-Giemsa staining of bone marrow of AML1-ETO9a-driven AML mice. (H–J) Percentage of GFP+ AML1-ETO9a AML cells in the (H) peripheral blood (PB), (I) bone marrow (BM), and (J) spleen (SP) of the mice treatment with DMSO or FB23-2 by flow cytometric analysis. (K–M) Flow cytometric analysis of the distribution of anti-CD11b-stained GFP + AML1-ETO9a AML cells in PB (K), BM (L), and SP (M) of mice treatment with DMSO or FB23-2. Fig. S4. Suppression of FTO resensitizes resistant cells to Ara-C in vitro and in vivo. (A and B) Apoptosis measured by flow cytometry for SKNO-1 (A) and Kasumi-1 (B) cells treated with DMSO, 30 µM Ara-C alone, 10 µM FB23-2 alone or combination of Ara-C and FB23-2 for 48 h with FB23-2 pretreatment for 6 h. (C and D) Percentage of GFP+ AML cells in peripheral blood (C) and spleen (D) of NOD/SCID/γcnull immunodeficient mice injected with SKNO-1 cells through tail vein treated with DMSO, Ara-C, FB23-2, or a combination of Ara-C and FB23-2 (n = 6 for each group). Fig. S5. Transcriptome-wide identification of FTO targets in t(8;21) AML. (A and B) Proportion of the distribution of m6A peaks in exon, intron, and intergenic regions across entire mRNA transcripts (A) or in the 5′-UTR, first exon, other exon, and 3′-UTR of mRNA transcripts (B) detected by m6A-seq assays in Kasumi-1 cells transduced with wild-type FTO or empty vector. (C and D) Gene set enrichment analysis (GSEA) of genes with a significant decrease in m6A levels as well as a significant increase (Hypo-up) or decrease (Hypo-down) in overall transcript levels in FTO-overexpressing Kasumi-1 cells. (E) Comparison of IGFBP2 expression between human primary AML cases with t(8;21) (n = 30, data from GSE30285) or normal controls (NC) (n = 9, data from GSE34814). (F) Kaplan–Meier analysis of event-free survival (left) and overall survival (right) of patients with AML (n = 344, data from GSE6891) based on the expression of IGFBP2. (G) Pearson correlation of the expression of FTO and IGFBP2 in BM samples of patients with AML from TCGA database (n = 173). Fig. S6. Luciferase reporter construction and identification of specific m6A readers targeting the 3′-UTR of IGFBP2 mRNA. (A) Construction of luciferase reporter vectors. Synthesized wildtype (wt) or mutant (mut) 3′ coding sequences of IGFBP2 were inserted into the XhoI and NotI site of the psiCHECK2 luciferase reporter. Putative m6A consensus motifs are shown in bold, whereas mutation sites (A to T mutation) are shown in red. (B and C) Identification of m6A specific binding proteins on 3′-UTR of IGFBP2 by RNA pull-down using 4 pairs of single-stranded RNA (ssRNA) baits containing the 4 m6A consensus motif on the 3′-UTR sequence of IGFBP2 respectively, with methylated (green) or unmethylated (red) adenosine (B). The iBAQ value of previously reported m6A readers (including YTHDFs, YTHDCs, IG2FBPs and hnRNPs) enriched by the 4 pairs of ssRNA probes detected by mass spectrometry analysis are shown (C). The YTHDC1, YTHDC3, and IGF2BP1 proteins that could not be enriched by all 4 pairs of ssRNA probes are not shown in the Figure (see Additional file 3: Table S4). (D and E) Western blot analysis of the expression of IGFBP2 with or without silencing of YTHDF2 (D) or YTHDF3 (E) by siRNA in SKNO-1 or Kasumi-1 cells. siNS, scramble siRNA. Fig. S7. Functional role of IGFBP2 in t(8;21) AML. (A and B) Effects of IGFBP2 knockdown on colony-forming capacity (A) and cell cycle (B) in SKNO-1 and Kasumi-1 cells. (C) Western blot analysis of silencing IGFBP2 by siRNA in SKNO-1 and Kasumi-1 cells. siNS, scramble siRNA. (D and E) External views (D) and weight (E) of the spleens from AML1-ETO9a-driven AML mice with or without Igfbp2 knockdown (n = 6 for each group) 7 weeks after transplantation. Fig. S8. FTO regulates leukemogenesis and sensitivity of t(8;21) AML cells to Ara-C through IGFBP2. (A) Effects of FTO knockdown with IGFBP2 overexpression after Ara-C treatment on colony-forming capacity of SKNO-1 and Kasumi-1 cells. (B–D) External views (B), weight of the spleens (C) and flow cytometric analysis of CD11b+ AML cells in PB, BM, and SP (D) of AML1-ETO9a-driven AML mice in Fig. 8E–G

A Carrier-Free Nanostructure Based on Platinum(IV) Prodrug Enhances Cellular Uptake and Cytotoxicity

Flurbiprofen, a hydrophobic COX inhibitor, was coordinated axially with oxoplatin to form a new conjugate, <i>cis</i>,<i>cis</i>,<i>trans</i>-[Pt­(IV)­(NH₃)₂Cl₂(flurbiprofen)₂]. The successful synthesis of this new conjugate was confirmed by ¹H, ¹³C, and ¹⁹⁵Pt NMR. The potential of this conjugate being reduced to cisplatin and subsequently exerting its DNA cross-linking ability was verified using cyclic voltammetry (CV), HPLC, and mass spectrometry (MS). This conjugate showed markedly higher cytotoxicity on many cancer cell lines than cisplatin, flurbiprofen, and their physical mixture (mole ratio, cisplatin:flurbiprofen = 1:2). This is consistent with the result of an apoptosis-inducing assay. This conjugate spontaneously assembles carrier-free nanoparticles in aqueous solution, which is confirmed by DLS, TEM, SEM, and AFM, and thus facilitates cellular uptake and markedly improves its cytotoxicity and apoptosis-inducing ability <i>in vitro</i>

Table_1_Perceptions of the benefits of the basic medical insurance system among the insured: a mixed methods research of a northern city in China.DOC

BackgroundThe perceptions of the benefits of the basic medical insurance system among the insured not only reflect the system's performance but also the public's basic medical insurance policy literacy, valuable information for countries that have entered the stage of deepening reform. This study aims to examine the factors that affect the perceptions of the benefits of the basic medical insurance system in China, diagnose the key problems, and propose corresponding measures for improvement.MethodsA mixed method design was used. Data for the quantitative study were obtained from a cross-sectional questionnaire survey (n = 1,045) of residents of Harbin who had enrolled for basic medical insurance system. A quota sampling method was further adopted. A multivariate logistic regression model was then employed to identify the factors influencing the perceptions of the benefits of the basic medical insurance system, followed by semi-structured interviews with 30 conveniently selected key informants. Interpretative phenomenological analysis was used to analyze the interview data.ResultsApproximately 44% of insured persons reported low perceptions of benefits. The logistic regression model showed that low perceptions of the benefits of the basic medical insurance system was positively correlated with the experience of daily drug purchases (OR = 1.967), perceptions of recognition with basic medical insurance system (OR = 1.948), perceptions of the financial burden of participation costs (OR = 1.887), perceptions of the convenience of using basic medical insurance for medical treatment (OR = 1.770), perceptions of the financial burden of daily drug purchases costs (OR = 1.721), perceptions of the financial burden of hospitalization costs (OR = 1.570), and type of basic medical insurance system (OR = 1.456). The results of the qualitative analysis showed that the key problem areas of perceptions of the benefits of the basic medical insurance system were: (I) system design of basic medical insurance; (II) intuitive cognition of the insured; (III) rational cognition of the insured; and (IV) the system environment.ConclusionsImproving the perceptions of the benefits of the basic medical insurance system of the insured requires joint efforts in improving system design and implementation, exploring effective publicity methods of basic medical insurance system information, supporting public policy literacy, and promoting the health system environment.</p

Table_1_Higher maternal thyroid resistance indices were associated with increased neonatal thyroid-stimulating hormone— analyses based on the Huizhou mother-infant cohort.docx

ObjectivesThis study aimed to explore the relationship of maternal thyroid function and thyroid resistance parameters with neonatal thyroid-stimulating hormone (TSH).MethodsThis work was a longitudinal study. Singleton pregnant women without a history of thyroid disorders were recruited in their first prenatal visit from October 2018 to June 2020. Maternal thyroid markers including TSH, free triiodothyronine (FT3), free thyroxine (FT4), and neonatal TSH were tested in the clinical laboratory of the hospital by electrochemiluminescence immunoassay. Thyroid resistance indices including Thyroid Feedback Quantile-based Index (TFQI), TSH index (TSHI), and thyrotroph T4 resistance index (TT4RI) were estimated in accordance with maternal FT4 and TSH levels. Multivariable linear and logistic regression was applied to explore the associations of maternal thyroid indices with infantile TSH level.ResultsA total of 3,210 mothers and 2,991 newborns with valid TSH data were included for analysis. Multivariable linear regression indicated that maternal thyroid variables were significantly and positively associated with neonatal TSH levels with standardized coefficients of 0.085 for TSH, 0.102 for FT3, 0.100 for FT4, 0.076 for TSHI, 0.087 for TFQI, and 0.089 for TT4RI (all P trend = 0.025], TFQI (OR = 1.746, 95% CI: 1.005–3.034; Ptrend = 0.016), and TT4RI (OR = 1.730, 95% CI: 1.021–2.934; Ptrend = 0.030) were significantly associated with an increased risk of elevated neonatal TSH (>5 mIU/L) in a dose–response manner.ConclusionThe longitudinal data demonstrated that maternal thyroid resistance indices and thyroid hormones in the first half of gestation were positively associated with neonatal TSH levels. The findings offered an additionally practical recommendation to improve the current screening algorithms for congenital hypothyroidism.</p

Transferrin-Dressed Virus-like Ternary Nanoparticles with Aggregation-Induced Emission for Targeted Delivery and Rapid Cytosolic Release of siRNA

Viruses have evolved to be outstandingly efficient at gene delivery, but their use as vectors is limited by safety risks. Inspired by the structure of viruses, we constructed a virus-mimicking vector (denoted as TR4@siRNA@Tf NCs) with virus-like architecture and infection properties. Composed of a hydrophilic peptide, an aggregation-induced emission (AIE) luminogen, and a lipophilic tail, TR4 imitates the viral capsid and endows the vector with AIE properties as well as efficient siRNA compaction. The outer glycoprotein transferrin (Tf) mimics the viral envelope protein and endows the vector with reduced cytotoxicity as well as enhanced targeting capability. Because of the strong interaction between Tf and transferrin receptors on the cell surface, the Tf coating can accelerate the intracellular release of siRNA into the cytosol. Tf and TR4 are eventually cycled back to the cell membrane. Our results confirmed that the constructed siRNA@TR4@Tf NCs show a high siRNA silencing efficiency of 85% with significantly reduced cytotoxicity. These NCs have comparable transfection ability to natural viruses while avoiding the toxicity issues associated with typical nonviral vectors. Therefore, this proposed virus-like siRNA vector, which integrates the advantages of both viral and nonviral vectors, should find many potential applications in gene therapy

Near-Infrared Emission CuInS/ZnS Quantum Dots: All-in-One Theranostic Nanomedicines with Intrinsic Fluorescence/Photoacoustic Imaging for Tumor Phototherapy

Many theranostic nanomedicines (NMs) have been fabricated by packaging imaging and therapeutic moieties together. However, concerns about their potential architecture instability and pharmacokinetic complexity remain major obstacles to their clinical translation. Herein, we demonstrated the use of CuInS/ZnS quantum dots (ZCIS QDs) as “all-in-one” theranostic nanomedicines that possess intrinsic imaging and therapeutic capabilities within a well-defined nanostructure. ZCIS QDs were exploited for multispectral optical tomography (MSOT) imaging and synergistic PTT/PDT therapy. Due to the intrinsic fluorescence/MSOT imaging ability of the ZCIS QDs, their size-dependent distribution profiles were successfully visualized at tumor sites <i>in vivo</i>. Our results showed that the smaller nanomedicines (ZCIS NMs-25) have longer tumor retention times, higher tumor uptake, and deeper tumor penetration than the larger nanomedicines (ZCIS NMs-80). The ability of ZCIS QDs to mediate photoinduced tumor ablation was also explored. Our results verified that under a single 660 nm laser irradiation, the ZCIS NMs had simultaneous inherent photothermal and photodynamic effects, resulting in high therapy efficacy against tumors. In summary, the ZCIS QDs as “all-in-one” versatile nanomedicines allow high therapeutic efficacy as well as noninvasively monitoring tumor site localization profiles by imaging techniques and thus hold great potential as precision theranostic nanomedicines