bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intracellular Expression of Neurotrophic Factors in Mononuclear Cells of Cord Blood and G-CSF-Mobilized Peripheral Blood As a Potential Source for Cellular Therapy
Authors: Lee Young-Ho , Koh Hani , Hwang Kyoujung 
Published: October 2019

Abstract
Purpose: To investigate a possible therapeutic mechanism for cell therapy in the field of neurological disorders using mobilized peripheral blood mononuclear cells (mPBMCs), we compared the expression of inflammatory cytokines and neurotrophic factors in PBMCs and mPBMCs from children with cerebral palsy (CP) to those from healthy adult donors and to cord blood (CB) donated from healthy newborns.
Methods: We evaluated the intracellular expression of neurotrophic factors and inflammatory cytokines in PBMCs and mPBMCs from 14 CP children and 14 healthy adult volunteer donors as well as CB mononuclear cells (CB-MNCs) donated from healthy newborns. For mPBMC, granulocyte colony stimulating factor (G-CSF, 10μg/kg/day) was administered subcutaneously for five consecutive days, and apheresis was performed on the fifth day to collect mPBMC via a central venous catheter. Both PBMC collected prior to G-CSF administration and apheresed mPBMC were cryopreserved. We performed flow cytometric analysis with intracellular staining for various cytokines after thawing PBMCs and mPBMCs as well as CBs.
Results: No significant differences in expression of neurotrophic factors were found between PBMC and mPBMC. However, in cells from CP children, the expression of IL-6 was significantly increased in mPBMC as compared to PBMC, and IL-3 was significantly decreased in mPBMC as compared to PBMC. In healthy adults, the expression of both IL-1β and IL-6 were significantly increased in mPBMC as compared to PBMC. The expression of BDNF in mPBMC from CP children was significantly higher than in CB or mPBMC from healthy adults. The expression of G-CSF in mPBMC from CP children was comparable to that in CB but significantly higher than in mPBMC from healthy adults. Lower expression of IL-1β, IL-3, and IL-6 and higher expression of IL-8 and IL-9 was observed from CB and mPBMCs from CP children rather than in healthy adults. Lower expression of IL-1β, and higher expression of IL-8 was observed from mPBMCs from CP children rather than in healthy adults.
Conclusion: The altered expression of neurotrophic factors and anti-inflammatory cytokines in mPBMC in CP children and CB from healthy children could provide a new potential source for cellular therapy for individuals with neurologic diseases.

Disclosures
Lee: Korea Healthcare Technology R&D Project (A101712), Ministry for Health & Welfare, Republic of Korea: Research Funding.
Study on Leukemia Cell Differentiation and Drug Resistance By SATB1 Gene
Authors: Wang Chunyan , Tan Huo , Lei Liu , Xu Lihua 
Published: October 2019

Abstract
Background and objective:
In this article, four types of lymphoma cell lines U937, Raji, Hut-102, Akata, and three types of leukemia cell lines K562, HL-60, Jurkat were used the expression of SATB1 in leukemia cells, Through cell differentiation induced by all-TRANS Retinoic acid and DMSO, finds the SATB1 gene expression is reduced, suggest that the SATB1 gene may be involved in cell differentiation. By gene silencing, reduce the SATB1 gene expression, sensitivity to chemotherapy drugs is increased, suggest that the SATB1 gene may be associated with drug resistance.
Methods:
1. Western Blot Assay for detection of SATB1 gene expression in cell lines.
2. DMSO and ATRA to HL-60, Jurkat cells for different times, SATB1 gene expression in the cells are detected by Western Blot.
3. Building SATB1-shRNA, transfected Jurkat cells and to HL-60.
4. Testing inhibition rate of chemotherapy drugs on HL-60, HL-60-SATB1-ctr, HL-60-SATB1-sh, Jurkat, Jurkat-SATB1-ctr, Jurkat-SATB1-sh cell, suggesting its relationship with the drug resistance.
Results:
1. SATB1 gene was expression in all the plant cells.
2. The expression of SATB1 in U937, Raji, Hut-102, Akata four types of lymphoma cell line is low.
3. It was highly expression in HL-60, Jurkat cells, but low in K562.
4. DMSO and ATRA treat HL-60, Jurkat cells for 48 and 96 hours, HL-60 cells become larger, rounded, Jurkat cells into smaller, is more obvious for 96-hour.
5. For HL-60, Jurkat cells, after DMSO and ATRA treated, the expression of SATB1 was decreased, more obvious for 96-hour.
6. HL-60-SATB1-sh1, Jurkat-SATB1-sh1 cell compared with the control group, the SATB1 protein content decreased significantly, successfully building SATB1-shRNA HL-60 and Jurkat cell line.
7. HL-60-SATB1-sh1, and Jurkat-SATB1-sh1 of daunorubicin (DNR) sensitivity has improved significantly.
Conclusion:
1. The SATB1 gene expression in leukemia and lymphoma cells, and higher expression in leukemia cells.
2. After DMSO and ATRA treatment, the SATB1 expression significantly reduced, suggest that SATB1 may be involved in cell differentiation.
3. Successfully built SATB1-shRNA, and successfully transferred to HL-60 and Jurkat cell lines.
4. HL-60-SATB1-sh1, and Jurkat-SATB1-sh1 of daunorubicin (DNR) sensitivity increased significantly, prompting SATB1 may be related to drug-resistance.

Disclosures
No relevant conflicts of interest to declare.
Decitabine Act As Demethylation Modulators in Acute Myeloid Leukemia for Reversal of Drug Resistance
Authors: Wang Zhixiang , Jiang Xuejie , Huang Kaikai , Yin Changxin , Zhong Qingxiu , Zhu Qiuhua , Ding Bingjie , Meng Fan Yi 
Published: October 2019

Abstract
Background and objectives The DNA methyltransferase inhibitors decitabine represent archetypal drugs for epigenetic cancer therapy. One alternative approach for the treatment of (Acute Myelocytic Leukemia) AML is the use of hypomethylating agents, including the 5-aza-2-deoxycytidine (decitabine; DAC) and other epigenetic regulators. However, the exact mechanism of DAC for drug resistance AML remains poorly understood. We conducted this study to explore how DAC regulated the sensitivity to adriamycin and aclacinomycin of AML cell line HL60/ADR and assessed the efficacy and safety of decitabine-based induction treatment patients with refractory acute myeloid leukemia.
Methods In this study, we analyzing cell proliferation of HL60, HL60/ADR and KG1-¦Á cells treated by DAC against control group detected with MTS assay. Then the changes of genes methylation were determined using Methylation specific PCR (MSP), stained with anti-CD11b, before being analyzed for cell markers with flow cytometric analysis. The cell protein levels were checked with Western blotting as described elsewhere. Data were collected from refractory acute myelogenous leukemia (AML) patients treated with homoharringtonine, cytarabine, and aclarubicin after decitabine (D-HAA). Control group consisted of fludarabine, cytarabine with or without Granulocyte Colony-Stimulating Factor between 2012 and 2014.
Results Different dose of DAC had proliferative inhibition effect on HL-60 cells, with time-dose dependent relationship. But the proliferative inhibition of DAC in KG1-¦Á and HL-60/ADR cells was not so effective. After been treated with 1¦ÌM DAC for 72 hours, HL-60/ADR cells were cultured with 0.5µg/ml adriamycin 1 hour. Adriamycin positive cells in DAC group were higher than control group, and the proliferative inhibition effect of adriamycin (P<0.001) and aclacinomycin (P<0.001) on HL60/ADR cells was increased (Figure 1). 1¦ÌM DAC significantly induced expression of CD11b on HL-60/ADR and KG1-¦Á cells, accompanied by cellular changes of differentiation (decreased nuclear¨Ccytoplasmic ratio, nuclear condensation, segmentation or lobulation, cytoplasmic granulation or vacuolization). Followed the increased the sensitivity of adriamycin, DNMT1 and SPRPs gene methylation was down-regulated. And ¦Â-catenin protein in nuclear was depressed after treatment of DAC. We enrolled 75 patients, of whom 71 were included in the intention-to-treat analysis. 65.2% patients in the D-HAA group achieved complete remission versus 29.2% in the FA/FLAG group (P=0.031). 2-year disease-free survival and overall survival were better in D-HAA group than FA/FLAG group (P=0.020, P=0.025; Figure 2).
Conclusion These ﬁndings provide evidence for clinic protocols using DAC to reversed drug resistance in refractory AML cell.

Disclosures
No relevant conflicts of interest to declare.
Fev Expression Is Functionally Required for Fetal HSCs and LSCs Indicating Prenatal Origin of Leukemia
Authors: Hong Dengli , Liu Tianhui , Zhang Fang , Liu Feng , Chen Guo-Qiang 
Published: October 2019

Abstract
Cancers, particularly in the childhood and adolescence, can originate prenatally or postnatally. How to distinguish however and what clinical implication of the distinct origin of cancers remains unknown. Here we have identified and characterized transcription factor fev an indicator of prenatal origin of leukemia in the human context of normal and leukemic hematopoiesis. Fev is expressed in fetal and neonatal and not in adult hematopoietic cells. Functional assays showed fev is essential for self-renewal in fetal HSCs. More interestingly, fev is expressed in leukemic cells of some cell lines and patients, which is associated with more frequent relapse. Fev-knockdown in leukemic cells markedly compromised their capacity of leukemic reconstitution in xenograft recipients. Leukemogenic targeting experiments by forced expression of MLL-AF9 or N-Myc showed fev-regulated self-renewal is retained from fetal HSCs rather than reactivated in adult HSCs. Therefore, fev-expression can distinguish LSCs from normal HSCs and indicate its prenatal origin.

Disclosures
No relevant conflicts of interest to declare.
Remission of a Heavily Pre-Treated Relapsed/Refractory ALK-Positive Anaplastic Large Cell Lymphoma after Treatment with Metronomic Therapy
Authors: Shunzong Yuan , Brad Haverkos , Su Hang , Yu Jianhua 
Published: October 2019

Abstract
The prognosis of ALK-positive, anaplastic large cell lymphoma is usually favorable, but patients that are resistant to primary chemotherapy or who relapse early have always been associated with an aggressive clinical course and unfavorable prognosis. For a recurrent relapsing case of ALK-positive, anaplastic large cell lymphoma heavily pretreated by nearly 50 cycles of chemotherapeutic agents (Fig 1.), we developed a treatment schema of metronomically scheduled low-dose methotrexate, which successfully controlled the relapse more than 12 months (Fig. 2). The metronomic therapy that we identified from this case was an effective and economic option that caused less damage to other organs than other treatments; thus, it should be given consideration for the treatment of similar cases, especially in the setting of where the recurrence seems to be only skin disease or when other effective regimens are unavailable. Our study represents the first report of a well-documented case of ALK+ALCL which was refractory to various lines of therapy but which was subsequently successfully treated with a combination of prednisone and etoposide followed by low-dose MTX maintenance metronomic therapy.
Figure 1 Figure 1.
Figure 2 Figure 2.

Disclosures
No relevant conflicts of interest to declare.
Platelets from Patients with Myocardial Infarction Can Activate T Cells in Vitro
Authors: Solomou Elena E. , Gizas Vasilios , Babali Theodora , Perperis Angelos , Verigou Evgenia , Ntavlouros Periklis , Gogos Charalambos , Alexopoulos Dimitrios 
Published: October 2019

Abstract
Specific aim: Our aim was to examine whether T cells can be activated by the circulating activated platelets from patients with myocardial infarction (with ST elevation-STEMI)
Methods: After written informed consent was obtained, peripheral blood mononuclear cells (PBMCs) were isolated from heparinized venous blood obtained from 20 patients with STEMI (18 men and 2 women) at the time of hospital admission at diagnosis, before receiving any treatment, as well as 5 days and 30 days later. We also analyzed 10 healthy subjects (8 men and 2 women), and 5 patients with unstable angina who served as the disease control group. PBMCs were analyzed by flow cytometry with the following markers and their isotypic controls: CD4, CD25, CD69, and FOXP3. We also isolated platelet rich plasma or plasma alone from the patients and the healthy subjects, and used in mixed cultures with PBMCs.
Results: We first examined T cell activation by measuring CD69 expression on CD4 T cells following incubation with platelets obtained from patients with STEMI. T cells treated with platelets from patients with STEMI showed increased expression of CD69 (as an activation marker) compared with T cells treated with platelets from healthy subjects (p<0.05, Figure 1). There was no T cell activation following incubation with plasma alone from patients or healthy controls.
We then examined the percentages of CD4+CD25+hi (regulatory T cells). There was no statistical difference in Tregs between patients at presentation and controls (healthy subjects and disease control group). Five days later, patients with STEMI displayed increased levels of Tregs compared with the 2 control groups; one month later, Treg numbers returned to the initial presentation levels (p<0.05, Figure 2)
Conclusion: To our knowledge we describe for the first time that platelets from patients with STEMI can activate T cells in vitro. In patients with STEMI, an increase in Tregs possibly in an effort to suppress immune system activation secondary to platelet activation, appears shortly after the infarct and normalizes a month later.
Figure 1. T cell activation after treatment with platelets from patients with STEMI compared with the platelets from healthy controls or plasma alone Figure 1. T cell activation after treatment with platelets from patients with STEMI compared with the platelets from healthy controls or plasma alone Figure 2. Increased T regs in patients with STEMI , 5 days after admission (STEMI EXIT) ( UA;unstable angina, STEMI 1mfup; STEMI after one month) Figure 2. Increased T regs in patients with STEMI , 5 days after admission (STEMI EXIT) ( UA;unstable angina, STEMI 1mfup; STEMI after one month)

Disclosures
No relevant conflicts of interest to declare.
Lower Dosages of Intravenous Immunoglobulin (IVIg) in Treating Immune Thrombocytopenia with Long-Term Follow-up of Three Years
Authors: Zhou Zeping , Qiao Zhuoqing , Li Huiyuan , Zhang Xian , Xue Feng , Yang Renchi 
Published: October 2019

Abstract

This study compared the effects of different dosages of intravenous immunoglobulin (IVIg) against immune thrombocytopenia. A total of 167 patients, 91 adults and 76 children, with ITP, followed-up for 3 years in the case-control study, were each divided into three subgroups according to the dosages of IVIg administered: group A (0.2g/kg/day), group B (0.3g/kg/day), group C (0.4g/kg/day). The therapeutic response in 91 adult patients did not differ significantly among the three groups of IVIg dosages (P=0.459). The response rate of IVIg treatment in the three adult groups was 97.1% for group A, and 97.2% for group B, 100% for group C. The mean time for raising platelets to 30 ×109/L in group A was 2.5 days, group B 3.2 days, group C 2.9 days (P=0.324). The median IVIg consumption in group A was 0.83 g/kg, group B 1.22 g/kg, and group C 1.64 g/kg (P<0.01). Similar results were shown in the children groups. The follow-up results showed no significant difference of clinical outcome between groups A, B and C. In conclusion, low-dose IVIg treatment is shown to be as effective as high-dose regimen without increasing the risk of developing the patients into chronic ITP conditions, suggesting that ITP patients could be treated more cost-effectively by lower than conventional dosage of IVIg regimen.

Disclosures
No relevant conflicts of interest to declare.
ONC201 Exhibits Mutation-Independent Efficacy with Superior Potency in Non-Hodgkin Lymphoma and Multiple Myeloma
Authors: Allen Joshua , Tarapore Rohinton , Garnett Mathew J , Benes Cyril 
Published: October 2019

Abstract
ONC201 is a first-in-class small molecule inducer of the integrated stress response that is currently in phase II clinical trials in select advanced cancers with promising early clinical results. The efficacy of this novel agent has been demonstrated in numerous preclinical advance cancer models in multiple indications with an exceptional safety profile that has translated well to the clinic. To determine the preclinical sensitivity profile of ONC201 in cancer, we performed an in vitro efficacy screen across >1,000 human cancer cell lines that represent a diverse array of tumor types and genetic aberrations. Sensitivity profiling was assessed by cell viability assays using dose responses curves at concentrations up to 20uM and at 72 hours post-treatment. Ranking the sensitivity dataset by tumor type, non-Hodgkin's lymphomas and multiple myeloma were the most sensitive tumor type to ONC201.
The mutation-agnostic efficacy that is most pronounced in lymphomas and multiple myeloma is in accordance with the recent findings that ONC201 induces the integrated stress response through a novel target to trigger is downstream late apoptotic effects. B-cell malignancies are particularly susceptible to induction of apoptosis via the integrated stress response, as they have relatively high basal activation of this pathway due to ER stress conferred by immunoglobulin production. Confirmatory studies revealed that multiple myeloma cell lines indeed possess pronounced sensitivity with nanomolar GI50s, unlike most other tumor types, that is particularly encouraging given the systemic concentrations observed in the first-in-man study. Together, these studies suggest specific advanced cancer indications, such as non-Hodgkin's lymphoma and multiple myeloma, as promising lead indications for this novel agent that are being evaluated in phase II clinical trials.

Disclosures
Allen: Oncoceutics, Inc: Employment, Equity Ownership. Tarapore:Oncoceutics, Inc: Employment, Equity Ownership.
Phase I Study of Single Agent Ibrutinib in Recurrent/Refractory Primary and Secondary CNS Lymphoma
Authors: Christian Grommes , Thomas Kaley , Abdel-Wahab Omar , Omuro Antonio M. , Ingo Mellinghoff , DeAngelis Lisa 
Published: October 2019

Abstract
BACKGROUND: Primary CNS Lymphoma (PCNSL) is an aggressive primary brain tumor. Standard treatment of PCNSL may include radiation, methotrexate-based therapy, and anti-CD20 antibody therapy (rituximab) and is associated with substantial morbidity and treatment recurrence. Outcome and treatment options for patients with recurrent/refractory disease are poor. There is only limited use of targeted agents in this patient population. Ibrutinib has shown promising clinical response in some B-cell malignancies. This phase I trial investigates the maximal tolerated dose of ibrutinib in patients with recurrent/refractory PCNSL and secondary CNS lymphoma (SCNSL).
METHODS: Eligible patients had a recurrent or refractory PCNSL or SCNSL, age≥18, KPS≥50, normal end-organ function, and unrestricted number and type of prior therapies. In patients with SCNSL disease, systemic disease needed to be absent or not requiring any active treatment.
RESULTS: Three patients have been enrolled at dose level 1 (560 mg daily) and one patient at dose level 2 (840mg daily) of whom three were women with a median age of 70 years (range 21-80). Three had recurrent PCNSL and 1 recurrent SCNSL. Two patients presented with parenchymal and two with leptomeningeal relapse. Treatment was generally well tolerated. There was one drug-related grade 4 toxicity (neutropenia) that resolved after the drug was held for 4 days. No drug related grade 3 toxicities have been observed to date. Most common grade 2 toxicities were decreased neutrophil count and hyperglycemia. All patients continue on study at a median follow up of 87 days. Three out of four patient were evaluated for response so far. There were two responses: one complete (in the CSF) and one partial, both in recurrent/refractory PCNSL as well as one stable disease in the patient with recurrent SCNSL. The patient with partial response had failed multiple prior treatment regimens including methotrexate-based chemotherapy, radiation, and rituximab/temozolomide. Serum and CSF pharmacokinetic analysis is initiated. Dose level 2 (840mg) is accruing.
CONCLUSION: Patients with CNS lymphoma tolerate Ibrutinib at 560 and 840mg well. Dose escalation will continue. Targeted agents might be an alternative therapeutic approach to be investigated for refractory/recurrent CNS lymphoma patients.

Disclosures
No relevant conflicts of interest to declare.
Evaluation of Modified Japanese Ministry Health and Labor Welfare Diagnostic Criteria for DIC to Detect Early Phase of DIC
Authors: Wada Hideo , Takeshi Matsumoto , Ohishi Kohshi , Katayama Naoyuki 
Published: October 2019

Abstract
Evaluation of the modified Japanese Ministry Health, Labour and Welfare (JMHLW) diagnostic criteria for DIC to detect the early phase of DIC
Introduction: Disseminated intravascular coagulation (DIC) is a syndrome characterized by the systemic activation of blood coagulation, which generates intravascular thrombin and fibrin, resulting in the thrombosis of small-to medium-sized vessels and ultimately organ dysfunction and severe bleeding. Three different diagnostic criteria incorporating similar global coagulation tests have been established by the International Society of Thrombosis and Haemostasis (ISTH), Japanese Ministry Health, Labour and Welfare (JMHLW) and Japanese Association of Acute Medicine (JAAM). The JMHLW score is well correlated with the severity of DIC and can be used to predict the outcome of the disease, however, it has a relatively low sensitivity for the early phase of DIC (pre-DIC).
Material and Methods: The Modified JMHLW criteria are shown in the Table 1. From 1996 to 2014, 282 patients with hematopoietic disorder type, 220 patients with infection type and 249 patients with other type were retrospectively diagnosed with DIC according to the JMHW criteria. There were 184 patients with pre-DIC and 256 patients without DIC.
Table 1. Modified Japanese Ministry Health, Labor and Welfare criteria for DIC Hematopoietic disorder type Othertype Infectiontype Platelet count(X107/ml) none 8 < < 12: 1 point, 5 < < 8: 2 points,(> 30% decrease: plus 1 point)< 5: 3 points FDP (ƒÊg/ml) 10 < < 20: 1 point, 20 < < 40; 2 points, 40< : 3 points Fibrinogen (g/L) 1 < < 1.5 : 1 point, < 1: 2 points none PT ratio 1.25 < < 1.67: 1 point, 1.67 < : 2 points AT (%) < 70 : 1 point TAT or SF 2X (upper limit of normal range): 1 point DIC 4 points = or < 6 points = or < 5 points = or < AT: antithrombin, TAT: thrombin AT complex, SF: soluble fibrin
Results: The agreement rate between the JMHLW and modified JMHLW criteria was 95.9%. The diagnosis rate of pre-DIC was 0% using the JMHLW criteira, 8.3% using the ISTH diagnostic criteria, 63.0% using the JAA criteria and 38.6% using the modified JMHLW criteira for DIC. The mortality of DIC was 42.5% using the JMHLW criteria, 45.5% using the ISTH criteria, 41.3% using the JAA criteria and 42.6% using the modified JMHLW criteria.
Discussion and conclusion: The agreement rate between the JMHLW and modified JMHLW criteria is high. As the modified JMHLW criteria adds the score for antithrombin (AT) activity, thrombin-AT complex (TAT) or soluble fibrin (SF), these criteria increase the sensitivity for pre-DIC.

Disclosures
No relevant conflicts of interest to declare.