[An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels].

To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Peking University People's Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients.

Curing hemophilia A by NHEJ-mediated ectopic F8 insertion in the mouse.

Background: Hemophilia A, a bleeding disorder resulting from F8 mutations, can only be cured by gene therapy. A promising strategy is CRISPR-Cas9-mediated precise insertion of F8 in hepatocytes at highly expressed gene loci, such as albumin (Alb). Unfortunately, the precise in vivo integration efficiency of a long insert is very low (~ 0.

Targeting of apoptosis gene loci by reprogramming factors leads to selective eradication of leukemia cells.

Applying somatic cell reprogramming strategies in cancer cell biology is a powerful approach to analyze mechanisms of malignancy and develop new therapeutics. Here, we test whether leukemia cells can be reprogrammed in vivo using the canonical reprogramming transcription factors-Oct4, Sox2, Klf4, and c-Myc (termed as OSKM). Unexpectedly, we discover that OSKM can eradicate leukemia cells and dramatically improve survival of leukemia-bearing mice.

Impact of Fibronectin Knockout on Proliferation and Differentiation of Human Infrapatellar Fat Pad-Derived Stem Cells.

Fibronectin plays an essential role in tissue development and regeneration. However, the effects of fibronectin knockout (FN1-KO) on stem cells' proliferation and differentiation remain unknown. In this study, CRISPR/Cas9 generated FN1-KO in human infrapatellar fat pad-derived stem cells (IPFSCs) was evaluated for proliferation ability including cell cycle and surface markers as well as stemness gene expression and for differentiation capacity including chondrogenic and adipogenic differentiation.

[Expression and influencing factors of hepcidin in classical paroxysmal nocturnal hemoglobinuria].

To investigate the serum expression and influencing factors of hepcidinin patients with classical paroxysmal nocturnal hemoglobinuria (PNH) . Retrospective analysis of 36 classical PNH patients from 2016.3 to 2017.

From organisational support to second victim-related distress: Role of patient safety culture.

Aim: This cross-sectional correlation study aimed to explore the associations among patient safety culture, organisational support, second victim-related distress, absenteeism and turnover intentions in the registered nurses (RNs) by using structural equation modelling.

Background: Medical errors could cause RNs' second victim-related distress, which may either positively or negatively impact patient safety culture.

Method: The Hospital Survey on Patient Safety Culture (HSOPSC) and Second Victim Experience and Support Tool (SVEST) were used to investigate 267 RNs who were involved in direct patient care and medical errors within the previous year in China.

[Long-term clinical effect of the CCLG-ALL2008 regimen in treatment of childhood acute lymphoblastic leukemia with different molecular biological features].

Objective: To study the long-term clinical effect of the CCLG-ALL2008 regimen in the treatment of children newly diagnosed with acute lymphoblastic leukemia (ALL) with different molecular biological features.

Methods: A total of 940 children who were newly diagnosed with ALL were enrolled in this study. The children were treated with the CCLG-ALL2008 regimen.

Metagenomic next-generation sequencing diagnosis of peripheral pulmonary infectious lesions through virtual navigation, radial EBUS, ultrathin bronchoscopy, and ROSE.

Objective: To evaluate the efficacy of combined rapid on-site evaluation of cytology (ROSE), ultrathin bronchoscopy, virtual bronchoscopic navigation, radial endobronchial ultrasound (EBUS), and metagenomic next-generation sequencing (mNGS) for diagnosis of peripheral pulmonary infectious lesions.

Methods: Specimens from patients with peripheral lung infection were obtained by transbronchial lung biopsy (TBLB) and bronchoalveolar lavage (BAL), and mNGS was used to detect pathogenic microorganisms. The sensitivity and specificity of mNGS were compared between TBLB tissue and BAL fluid.

Liver Kinase B1 Fine-Tunes Lineage Commitment of Human Fetal Synovium-Derived Stem Cells.

Liver kinase B1 (LKB1), a serine/threonine protein, is a key regulator in stem cell function and energy metabolism. Herein, we describe the role of LKB1 in modulating the differentiation of synovium-derived stem cells (SDSCs) toward chondrogenic, adipogenic, and osteogenic lineages. Human fetal SDSCs were transduced with CRISPR associated protein 9 (Cas9)-single-guide RNA vectors to knockout or lentiviral vectors to overexpress the LKB1 gene.

Nrf2/Keap1/ARE Signaling Mediated an Antioxidative Protection of Human Placental Mesenchymal Stem Cells of Fetal Origin in Alveolar Epithelial Cells.

The oxidative stresses are a major insult in pulmonary injury such as acute lung injury (ALI) and acute respiratory distress syndrome (ARDS), two clinical manifestations of acute respiratory failure with substantially high morbidity and mortality. Mesenchymal stem cells (MSCs) hold a promise in treatments of many human diseases, mainly owing to their capacities of immunoregulation and antioxidative activity. The strong immunoregulatory role of human placental MSCs of fetal origin (hfPMSCs) has been previously demonstrated; their antioxidant activity, however, has yet been interrogated.

[Dynamics of Proliferation and Differentiation after Transplantation of Hematopoietic Cells in Mouse].

Objective: To observe the dynamic changes of hematopoietic reconstitution and multiple lineages differentiation at early phase after transplantation.

Methods: Whole bone marrow mononuclear cells (wBMMNC, 5×10) and enriched c-Kit hematopoietic stem/progenitor cells (HSPC, 3×10) from the BM of B6-Ly5.1 mice were transplanted into lethally irradiated B6-Ly5.

[Comparison of Effect of Serum-Free Culture Systems on Directional Erythroid Differentiation of Human Umbilical Cord Blood CD34 Cells].

Objective: To compare the efficacy of directional erythroid differentiation in different serum free culture systems and to screen the optimal culture systems for inducing the differentiation of umbilical cord blood hematopoietic stem and progenior cells (HSPC) to erythroid cells.

Methods: The CD34 cells from umbilical blood munonuclear cells were sorted by using the magnetic beads, and were inoculated into 3 different of culture systems (system 1, 2 and 3 respectively), to induce erythrold differentiation by 3 stage culture. The living cells were counted in different differentiation stages and were observed by Wright-Giemsa staining; the expression of CD71 and CD235a on cell surface was detected by flow cytometry, the erythroid differentiation pteency was detected via colony-forming test.

[Clinical Features and Therapeutic Efficacy in Adult Acute Lymphoblastic Leukemia with t (1; 19) (E2A-PBX1)].

Objective: To explore the clinical features and therapeutic efficacy in adult ALL patients with t (1; 19) (E2A-PBX1).

Methods: The clinic data of 19 adult ALL patients with t (1; 19) (E2A-PBX1) in our hospital from Nov. 22, 2010 to Apr.

New paradigms on hematopoietic stem cell differentiation.

Ever since hematopoietic stem cells (HSCs) were first identified half a century ago, their differentiation roadmap has been extensively studied. The classical model of hematopoiesis has long held as a dogma that HSCs reside at the top of a hierarchy in which HSCs possess self-renewal capacity and can progressively give rise to all blood lineage cells. However, over the past several years, with advances in single cell technologies, this developmental scheme has been challenged.

PASS: A Proteomics Alternative Splicing Screening Pipeline.

Alternative splicing (AS) has been well-investigated at the trancriptome level by the application of RNA-seq technology. There is an ongoing debate on the biological importance of AS to proteome complexity. A toolkit for accurately identifying AS from proteome data is urgently needed.

USP9X-mediated deubiquitination of B-cell CLL/lymphoma 9 potentiates Wnt signaling and promotes breast carcinogenesis.

Hyperactivation of the canonical Wnt-signaling pathway is a prominent feature of a number of human malignancies. Transcriptional activation of this signaling cascade depends on the formation of the β-catenin-B-cell CLL/lymphoma 9 (BCL9)-pygopus (PYGO) family plant homeodomain finger 1 complex, yet how the assembly of this complex is regulated remains to be investigated. Here, using MCF-7, HeLa, HEK293T, MDA-MB-231, and Sf9 cells, along with immunoblotting and immunofluorescence, nano-HPLC-MS/MS, deubiquitination, immunoprecipitation, and chromatin immunoprecipitation (ChIP) assays, we report that BCL9 physically associates with a protein deubiquitinase, ubiquitin-specific peptidase 9, X-linked (USP9X), and that USP9X removes Lys-63-linked polyubiquitin on Lys-212 of BCL9.

[Role of Rictor in Hematopoietic Stem Cells during Fetal Liver Hematopoiesis].

Objective: To investigate the effect of Rictor on the hematopoiesis of fetal liver by specific knock-out of Rictor in hematopoietic cells of Vav-Cre mice.

Methods: E12.5 0.

[Screening and Verification of Antioxidant Small Molecular Compounds for Expansion of Human Hematopoietic Stem Cells Ex Vitro].

Objective: To screen the antioxidant small molecular compounds with optimal efficiency of expansing the human hematopoietic stem cells (hHSC) In vitro based on antioxidant small molecular compound database of LKT laboratory, and to verify the effects of these compounds on the biological functions of hHSC.

Methods: The umbilial cord blood CD34 cells were enriched by using the MACS beads; the absolute number and percentage of CD34 cells and CD34 CD49f cells were detected by high throughput flow cytometry after culture of hHSC with compounds in vitro for 1 week, the SR1 (1 μmol/L) was used as positive control, the candidate compounds were screened out; then 4 compounds were selected for follow-up experiments by comprehensive evaluation of concentration, safety and expansion efficacy, the optimal used concentrations of selected compounds were determined through the concentration gradient analysis, and CFC short-term colony-forming cell test was performed by using the determined concentration so as to verify the effect of compounds on the self-renewal, multilineage differentiation.

Results: Out of 85 antioxidant small molecular compounds, 4 compounds (C2968, D3331, B1753 and B3358) with obvious expansion efficacy for CD34 cells and CD34 CD49f cells were screened out by high throughput flow cytometry; their optimal concentrations of 4 compounds were 0.

[Symptomatic Profiles of 173 Chinese Patients with Essential Thrombocythemia].

Objective: To explore the symptomatic burden of patients with essential thrombocythemia (ET) and its relation with clinical characteristics including the mutation status, therapeutic protocols and sex.

Methods: Total of 173 Chinese ET patients were selected and grouped on the basis of disease characteristics (mutation status, therapeutic pro to- cols, and sex).

Results: All the groups showed low-to-high symptom burden, with the highest in the Hu (hydroxyurea)-group (total symptom score [TSS], 14.

[Role of Vascular Niche of Bone Marrow in the Development of MLL-AF9 Induced Acute Myclaid Leukemia].

Objective: To investigate the role of bone marrow vascular niche in the development of MLL-AF9 acute myeloid leukemia (AML).

Methods: Transplantation experiments were performed to establish non-radiated MLL-AF9 AML model; the half-bone immunofluorescence staining and tile-scan imaging of two-photon confocal microscopy were used to obtain the data of 3 main bone marrow niche cells; flow cytometry analysis was performed to characterize leukemia cells in different anatomical sites.

Results: In the early stage of MLL-AF9 AML, the proportion of leukemia cells in the metaphysis of the femur was significantly higher than that in the diaphysis.

A new HLA-C allele, HLA-C*15:144, identified in a Chinese family.

HLA-C*15:144 differs from HLA-C*15:07 (343G->A, G91R).

Using local alignment to enhance single-cell bisulfite sequencing data efficiency.

Motivation: Single-cell bisulfite sequencing (BS-seq) techniques have been developed for DNA methylation heterogeneity detection and studies with limited materials. However, the data deficiency such as low read mapping ratio is still a critical issue.

Results: We comprehensively characterize single-cell BS-seq data and reveal chimerical molecules to be the major source of alignment failures.