bluuebunny/crossref_metadata_2025_split huggingface dataset:
RNA Sequencing Based Whole Transcriptome Analysis Detected Precisely All Fusion Transcripts in Leukemias
Authors: Shin Myung Geun , Lee Jun Hyung , Choi Hyun Jung , Kee Seung Jung , Kim Soo Hyun , Shin Jong Hee , Suh Soon Pal 
Published: October 2019

Abstract
Introduction: Fusion transcript is a chimeric RNA encoded by a fusion gene or by two different genes by subsequent trans-splicing. Detection of fusion transcripts is an integral part of routine diagnostics of hematological malignancies. However, most of previous analytical methods couldn't detect all fusion transcripts in leukemia. In this study, we developed accurate fusion transcript detection methood using whole transcriptome sequencing, fusion gene detection software and expression analysis.
Methods: RNA sequencing (RNA-seq) for whole transcriptome was performed in 11 patients with hematological malignancies (4 AML, 2 APL, 2 ALL, and 3 CML) having fusion transcripts detected by multiplex RT-PCR (HemaVision, DNA Diagnostic, Risskov, Denmark). Library were prepared with 1 ug of total RNA for each sample by TruSeq mRNA Sample Prep kit (Illumina, San Diego, USA). The libraries were quantified using qPCR according to the qPCR Quantification Protocol Guide (KAPA Library Quantificatoin kits for Illumina Sequecing platforms) and qualified using the TapeStation D1000 ScreenTape (Agilent Technologies, Santa Clara, USA). Indexed libraries were then sequenced using the HiSeq2500 platform (Illumina). The data obtained from the sequencing was analyzed using STAR-Fusion (v1.2.0). Novel fusion transcripts were confirmed by conventional sequencing.
Results: Using STAR-Fusion, average number of fusion candidates per sample was 949.8 (range, 286-1752). To exclude false positive results and obtain true positive results, we developed the following filtering algorithm. First filtering criterion is to have more than 5 junction reads, the second is to detect more than one number of spanning reads, and the third criterion is to be in-frame fusion, which type of fusion can actually synthesize intact protein. Fusion candidates remaining after applying the above three filtering criteria were 1-3 per sample. All known fusion transcripts (PML--RARA, RUNX--RUNX1T1, CBFB--MYH11, KMT2A--MLLT3, BCR--ABL1, DEK--NUP214, ETV6--RUNX1) by multiplex RT-PCR were also detected in RNA-seq. In addition, 10 novel fusion transcripts (IGKV4-1--IGKC, IGLV1-47--IGLC2, HBA2--HBB, DEFA3--MBNL1, HBB--HBA2, MPO--HBA2, HBS1L--AHI1, HBB--HBA2, IGKV4-1--IGKC, SS18L1--ADRM1) were detected and among them, 6 fusions were confirmed by conventional sequencing.
Conclusions: Whole transcriptome sequencing and optimized filtering algorithms successfully detected all known fusion transcripts and various novel fusions.

Disclosures
No relevant conflicts of interest to declare.
Decitabine Combined with FB3 As Conditional Regimen for Elderly Acute Myeloid Leukemia Patient with Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Gao Shichun , Liu Jia , Peiyan Kong , Gao Li , Zhang Cheng , Liu Yao , Yan Hongju , Zhu Lidan , Zeng Defu , Gao Lei , Zhang Xi 
Published: October 2019

Abstract
Background
Acute myeloid leukemia (AML) is a malignant hematologic disease with high incidence in the elderly peoples. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an important method for the treatment of AML. However, elderly patients with AML are unable to receive myeloablative conditioning, and transplant related mortality (TAM) is significantly higher than that of young AML patients. How to reduce TAM in elderly AML patients, improve the efficacy of allo-HSCT, and further expand the applicable population of allo-HSCT is the focus of clinical research.
Patients and methods
According to the clinical trial (ChiCTR-IIR-16008182), elderly patients with AML used Decitabine (Dec) combined with FB3 as conditional regimen (20mg/m2 decitabine on day -11∽-7, 30mg/m2 Fludarabine on day -6∽-2, 130mg/m2 busulfan on day -4∽-2). The primary outcome of the trial was relapse after transplantation, and the secondary outcomes were chronic graft versus host disease (cGVHD) and the safety of treatment.
Results
From Mar 2016 to Jun 2018, 19 elderly patients with AML in our center include in the study. The median age was 62 (60-66) years. According to cytogenetics and molecular mutation, 8 patients were intermediate risk and 11 patients were unfavorable risk. HLA matched HSCT was performed in 7 patients, and HLA haploidentical HSCT in 12 patients. All of the patients achieved complete donor chimerism by day 28 after HSCT. Patients achieved ANC engraftment at 15 (10-19) days and PLT engraftment at 19 (12-26) days. Five patients relapsed after transplantation, and 4 of them died; one patient died of severe acute graft versus host disease (aGVHD) and pulmonary infection. The main adverse reactions during the conditioning treatment were dizziness, nausea, vomiting, diarrhea and oral ulcers. ALL symptoms were relieved after the symptomatic treatment. Three patients had a mild elevated transaminase and improved after liver-protecting treatment. Till now, the leukemia-free survival and overall survival were 73.68%.
Conclusion
Dec combined with FB3 as conditional regimen was well tolerated and can achieve ideal survival. The multicenter, randomized controlled clinical study will conduct to further verify its effectiveness and safety.
Key Words: Decitabine, FB3, Elderly, Acute myeloid leukemia patient, Allogeneic hematopoietic stem cell transplantation

Disclosures
No relevant conflicts of interest to declare.
MDS/Secondary AML Associated with Type I Gaucher Disease
Authors: Okamoto Atsushi , Yamamoto Kenta , Eguchi Go , Kanai Yoshitaka , Yamaguchi Terufumi , Maeda Yasuhiro 
Published: October 2019

Abstract
[Introduction]
Gaucher disease is one of lysosomal diseases, and it is an extremely rare disease characterized by anemia, thrombocytopenia, splenomegaly, multiple pathologic fractures and so on. It has been reported that Gaucher disease often merges with blood malignant diseases. However, there are no reports of myelodysplastic syndrome/secondary AML (MDS/s-AML) concomitant with type I Gaucher disease. Furthermore, we carried out parallel treatment of enzyme replacement therapy for type I Gaucher disease and azacitidine therapy for MDS/s-AML.
[Case]
A 69-year-old woman admitted to our hospital because of multiple abnormal fractures. She was diagnosed of MDS (refractory anemia) in 2011 by a previous doctor. As anemia and thrombocytopenia gradually worsened on May 2017, she was transferred to our department on August 8, 2017. At the time of visit, typical pneumonia determined with chest X-P and CT has been observed and high value of CRP (11.0mg/dl) was existed. Pneumonia improved promptly with antibiotic therapy, however, high value of CRP continued, and bilateral chest and right back pain got worse. We carried out plain CT and MRI (DWIBS). In results, splenomegaly, bilateral rib fracture and osteolytic changes in the right iliac bone were observed. Although bone marrow biopsy was performed twice without significant findings, Gaucher disease has been suspected and glucocerebrosidase activity was measured. As expected, a significant decrease of the enzyme activity was observed. Finally, a diagnosis of type I Gaucher disease was made. We started enzyme replacement therapy for glucocerebrosidase. Therefore, reduction of splenomegaly and independency of transfusion were induced by several replacement therapy. Further, multiple bone fracture lesions have improved. Further, juvenile leukocyte including myeloblasts in peripheral blood (PB) also decreased. Therefore, we suspected that the replacement therapy improved not only several symptoms of Gaucher disease, but several hematological findings of MDS. However, WT1-mRNA in the peripheral blood has gradually increased. Moreover, progression of MDS/s-AML was observed after two months. We selected azacitidine therapy in considering of performance state of the patient. In addition to enzyme supplementation therapy, azacitidine therapy was performed in parallel. After several treatment of azacitidine, blasts in PB were decreased significantly with independency of transfusion.
[Conclusion]
We deeply indicate that the combination of enzyme replacement therapy and azacitidine therapy is useful for cases of MDS/s-AML associated with Gaucher disease. There is a possibility that the reduction of the enzyme is involved not only in Gaucher disease but in the ontogeny of blood diseases such as MDS. The effects of enzyme replacement therapy of Gaucher's disease for MDS/s-AML should be discussed.

Disclosures
No relevant conflicts of interest to declare.
Impact of High Donor-Derived CD4+ CCR7+ T Cell Count on Incidence of Grade II-IV Acute Graft-Versus-Host Disease Following Haploidentical Hematopoietic Cell Transplantation
Authors: Varlet Pauline , Alsuliman Tamim , Trauet Jacques , Demaret Julie , Labalette Myriam , Yakoub-Agha Ibrahim 
Published: October 2019

Abstract
Introduction
Historically the administration of post-transplant high-dose Cyclophosphamide (PTCy) has led to haplo-HCT re-innovation.
Although PTCy has a positive impact towards reducing severe acute graft-versus-host disease (aGVHD), this drug has serious adverse side effects and makes haploidentical-HCT more difficult in some patients-particularly in older patients and/or those presenting other comorbidities.
Considering our previous published experience in the effect of graft lymphocyte composition, this work aims to explore the impact of infused T cell subsets on Overall Survival (OS), Event Free Survival (EFS) and acute GVHD in a retrospective cohort receiving allogeneic haplo-HCT.
Methods
This study retrospectively analyzed 29 adult patients who underwent first allogeneic haplo-HCT for hematologic malignancies at Lille University Hospital (CHRU Lille, France).
Graft samples were analyzed by flow cytometry, and CD4 and CD8 T cell subsets were defined as follows: TN=naïve T cells; , TCM=central memory T cells; TEM=effector memory T cells; TTD =terminally differentiated T cells.
Results
The median follow-up of patients was 11 months (0.4-44.3). The cohort median recipient age was 59 years old. Cumulative incidences of grade 2 to 4 aGVHD were 31%. The rate of grade 3 to 4 severe GVHD was 17%. Eleven patients died with 14% of deaths due to non-relapse mortality.
We found a correlation between high percentage of donor-derived CD4+ CCR7+ T cells (>69.2% for CD4+ T cells-the median value in our study) and aGVHD (p=0.028) without any impact on OS and EFS (Table 1).
In multivariate analysis, only high proportions of donor CD4+ CCR7+ T cells correlated significantly with aGVHD (HR=0.203, 95% CI [0,042-0,980], p=0,047) without any impact on OS and EFS (Figure 1).
Conclusion
Naïve and central memory T cells expressing CCR7 exhibit higher alloreactivity potential than CCR7- T cells. In this study, even with PTCy administration, we observed that a high percentage of donor-derived CD4+ CCR7+ T cells can be considered as a predictive indicator of grade II-IV aGVHD post Haplo-HCT. Thus, selective depletion of CD4+ CCR7+ T cells might be enough to prevent aGVHD in haplo-HCT, enabling the use of low doses of PTCy in order to reduce post haplo-HCT complications.

Disclosures
No relevant conflicts of interest to declare.
The Effect of Ketamine Administration on Pain Control in Painful Crisis of Sickle Cell Anemia Patients during Childhood: A Retrospective Observational Study
Authors: Kilinc Yurdanur , Temizturk Hacer , Yılmaz Hayri levent , Cil Metin , Leblebisatan Goksel , Sasmaz Hatice Ilgen 
Published: October 2019

Abstract
Background: Sickle cell anemia (SCA) is a hemoglobinopathy as a result of substitution of glutamic acid with valine in the 6th position of the β globin chain. The vaso-occlusive crises of patients are often caused by pain that requires in-patient treatment. The attack rate is reported to changed between 0.5-1.0 attacks per year per patient. It could be reach up to 10% in 5% of patients. Identification of the SCA painful crisis and the determination of appropriate treatment is very important for the pain relief. Recently studies support that ketamine treatment is beneficial in patients who do not respond to traditional methods. Ketamine is a non-competitive antagonist at the NMDA receptor. We used retrospective visual analogue scale (VAS) scoring system in acute painful crisis patients and then applied ketamine and tramadol to investigate the effect of treatment and side effects and the maximum change in pain score within the first hour after drug administration retrospectively. The aim of this study is evaluate the level of ketamine effect in painful crises.
Methods: Patients older than 6 years of age with acute painful crisis, defined as VAS of greater or equal to 4 were enrolled.The retrospective data was collected from electronic records and included age, sex, vital signs, changes in pain score within 1 hour, and side effects. Ketamine (0.25 mg / kg / dose) and tramadol (0.1-0.4 mg / kg / h) were administered. Patients with a pain score of <4 or a decrease of ≥3 points in VAS were considered to be responsive.
Results: Ketamine group (n = 16), 6 female (37.5%), 10 male (63.5%) and the mean age was 15.4 (7-21) years. Tramadol group (n = 31) 15 female (48%), 16 male (52%) and the mean age was 15.6 (6-21) years. The age and sex groups were similar (p> 0.05). After drug administration, vital signs were observed in all patients in normal range to their age. In the tramadol and ketamine groups, the number of patients who responded at 20 minutes were 9(29%)-9(56%), 12(38%)-11(68%) (p<0.05) at 60 minutes. Sixteen patients (51.6%) responded in the 6th hour of the tramadol group, no evaluate in ketamin group. No life-threatening side effects was observed in any patient.
Conclusion: In the treatment of painful crises, ketamine administration was found to be more effective than tramadol in the first hour. Due to the longer duration of the initial effect of tramadol treatment, ketamine should be considered as an option in the rapid treatment of painful crises.

Disclosures
No relevant conflicts of interest to declare.
Long Term Outcome Following Splenectomy in Children
Authors: Akca Tugberk , Salcioglu Zafer , Aydogan Gonul , Tahtakesen Güçer Tuba Nur , Bayram Cengiz , Ozdemir Gul Nihal 
Published: October 2019

Abstract
Aim: The aim of the study is to evaluate the efficacy, complications and long term results of splenectomy in children with hematologic diseases.
Patients and Methods: This study includes retrospective analysis of the medical records of 158 children between 0-18 years old who underwent splenectomy due to hematologic diseases from 1989 to 2017. Gender, splenectomy indications, age of diagnosis, splenectomy age, vaccination status, splenectomy outcomes, complications and mortality were evaluated.
Results: Of the cases, 88 (55.7%) were female and 70 were male (44.3%). Indications for splenectomy were hereditary spherocytosis in 59 (37.3%), immune thrombocytopenia (ITP) in 50 (31.6%) and thalassemia major in 40 (25.3%) children. The median age of diagnosis was 3.9 years, the median splenectomy age was 9.2 years, the median time from diagnosis to splenectomy was 5.3 years in all cohort. Mean follow-up time of patients after splenectomy was 7.3±5.2 years. All patients except one were immunized before splenectomy. Responses were assessed based on standart criteria for each disease. Response was seen in 94.7% of patients with hereditary spherocytosis and 80% of ITP patients. Twenty-two percent of splenectomised children with thalassemia major had a decrease in transfusion frequency. Infections following splenectomy were reported in 22.8%. One patient had thrombosis related to splenectomy, none of the cases had major bleeding. There was no death due to sepsis. One patient with ITP died due to Subacute Sclerosing Panencephilitis (SSPE).
Conclusion: Splecetomy is associated with good remission rates following ITP and hereditary spherocytosis. Splenectomy should be performed only in selected cases with transfusion dependent thalassemia. Post-splenectomy sepsis is uncommon. Future studies should focus on health quality of life and cost analysis of splenectomy in children.

Disclosures
No relevant conflicts of interest to declare.
Differential Outcomes of Synchronous and Metachronous Diffuse Large B-Cell Lymphoma Transformed from Follicular Lymphoma
Authors: Chan Natalie Pei Xin , Lim Ryan Mao Heng , Khoo Lay Poh , Cheng Chee Leong , Tan Leonard Kwan Cheong , Poon Eileen Yi Ling , Somasundaram Nagavalli , Farid Mohamad , Tang Tiffany , Tao Miriam , Lim Soon Thye , Chan Jason Yongsheng 
Published: October 2019

Abstract
Aim:
Composite histologies of diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL) may be present synchronously at diagnosis or metachronously at the time of transformation of a formerly diagnosed FL. The aim of this study was to examine their clinico-pathological characteristics and treatment outcomes.
Method:
Patients who were consecutively diagnosed with composite FL/DLBCL (n=120) and FL (n=346) from 2001-2017 at the National Cancer Centre Singapore were retrospectively analyzed. Chi-squared tests and multivariate logistic regression were performed to evaluate clinico-pathological associations between the two cohorts. Survival analysis was performed using the Kaplan-Meier method and the log-rank test.
Results:
Amongst the FL cases, 21 patients (6.1%) with metachronous transformed FL/DLBCL were identified. The median lag time from diagnosis of FL to DLBCL transformation was 47 months (range, 7.8-168). Clinico-pathological features in synchronous and metachronous FL/DLBCL were similar, with both entities demonstrating a male preponderance (67% male and 33% female). Median age at diagnosis was 67 years (range, 41-81) and 60 years (range, 24-90) for metachronous and synchronous FL/DLBCL, respectively. The cell-of-origin by Han's criteria was similar (metachronous: GCB 52%, ABC 43%, unknown 5%; synchronous: GCB 38%, ABC 57%, unknown 5%; p = 0.21), as were the occurrence of C-MYC/BCL2/BCL6 double-hit rearrangements. However, survival from the time of DLBCL development was significantly worse (median, 3 vs 12 years) for metachronous compared to synchronous FL/DLBCL (HR 2.20, 95%CI 0.88-5.49, p = 0.022). Double-hit, advanced stage, and use of non-RCHOP regimens (OR 7.54, 95%CI 2.84-20.1, p = 0.0001) were associated with lack of complete response to chemotherapy. In metachronous FL/DLBCL, the R-CHOP regimen was less commonly used (77% vs 56%, p = 0.049). Correspondingly, complete response to chemotherapy was less likely in metachronous cases (38% vs 63%, p = 0.037).
Conclusion:
Metachronous and synchronous FL/DLBCL share similar clinico-pathological characteristics.
A preceding diagnosis of FL however, predicts for significantly worse survival outcomes and suboptimal responses to chemotherapy.

Disclosures
No relevant conflicts of interest to declare.
Cell-Free DNA Analysis Demonstrates Comprehensive Genomic Profiling and a Facile Method to Sensitively in Multiple Myeloma
Authors: Du Juan , Lu Jing , Qiang Wanting , Li Lu , Liu Jin , Jiang Hua , Qian Ziliang , Chen Baoan , Fu Weijun 
Published: October 2019

Abstract
Background: Multiple myeloma (MM) is a plasma cell malignancy characterized by complex cytogenetic and molecular abnormalities including translocations involving the immunoglobin heavy chain locus and mutations involving numerous oncogenic signaling pathways. Fluorescence in situ hybridization (FISH) has emerged as the most useful current cytogenetic assessment and provide a new level of insight into the correlation of myeloma prognosis risk model. However, the identification or sorting of malignant cells is required before FISH probes and only involved expansion of the types of probe and number of detectable targets is reached. Cell-free DNA (cfDNA) offers the potential for minimally invasive genome-wide profiling of tumor alterations without tumor biopsy and may be associated with cancer precision medicine and patient prognosis.
Methods: In this retrospective cohort study, we identified 37 patients from 9 relapsed/refractory (RR) and 33 newly diagnosed (ND) patients were analyzed for chromosomal copy number imbalance using the ultrasensitive chromosomal aneuploidy detector (UCAD) platform.
Results: Chromosome copy number aberration (CNA) were frequently (82.6%, N=46) detected in MM plasma cell free DNA. Applying UCAD to cfDNA, FISH in CD138 purified bone marrow aspirates, and some matched bone marrow biopsies, we find concordance in copy number alterations (~81%) between liquid and tumor biopsies. Significant copy number changes, including 1q gains, 13q deletion and 17p deletion could be found in 57.89%, 54.05%, and 16.67% in plasma of MM, which is higher percentage than FISH assay (46.81%, 28.26%, and 8.89%), respectively. Besides, chromosome 6p and 6q were determined the higher frequency aberration from UCAD. Moreover, a higher frequency of copy number aberrations and variations was detected in RR patients than ND (100% vs 78.4%, respectively), obviously CNAs heterogeneity displayed in advanced disease. In the inconsistent some samples, UCAD from the plasma and bone marrow showed the similar results, which indicated the FISH is underdetermined and insensitivity in some patients' routine inspection.
Conclusion: We conclude that cfDNA analysis as an adjunct to BM biopsy represents a noninvasive and broaden the applicability strategy for comprehensive genomic profiling and therapeutic monitoring of MM.

Disclosures
No relevant conflicts of interest to declare.