bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Effect of Phenylhexyl Isothiocyanate on Drug Resistance of K562/A02 Cell Line
Authors: Chen Bao-An , Yuan Peng , Cheng Jian , Gao Feng , Ding Jia-Hua , Gao Chong , Sun Yun-Yu , Wang Jun , Zhao Gang , Song Hui-hui , Bao Wen , Shao Ze-ye , Xia Guo-hua , Lu Quan-yi , Ma Xu-dong , Liu De-long , Wang Jue-qiong , Pei Xiao-ping , Chen Ning-na 
Published: April 2020

Abstract
Objective: To investigate the effect of 6-phenylhexyl isothiocyanate (PHI) on drug resistance and sensitivity on K562/A02 cell line to ADM and elucidate the probable mechanisms.
Methods: We measured growth inhibition of ADM on K562/A02 cell line by MTT assay. Apoptosis rate of K562/A02 cell line, the change of intracellular ADM and MRP1 protein level were detected by Flow Cytometry. Intracellular deoxidized GSH by spectrometric enzyme assay and MRP1 mRNA by RT-PCR semiquantitative assay were observed anteroposterior using PHI.
Results: PHI can enhance the sensitivity of K562/A02 cell line to ADM, survival rate of K562/A02 cell line decreased with the increasing concentration of PHI and ADM. Apoptosis rate increased with treating by combination of two above drugs, and multiple of drug resistance had statistical significance (P&lt;0.05) when the concentration of PHI was more than 20μmol/l. Intracellular GSH of K562/A02 cell line reduced 5% when 1μg/ml ADM was single used, and when more than 10μmol/l PHI was used it increased slightly at first then decreased. When more than 20μmol/l PHI and 1μg/ml ADM were used combination, intracellular GSH of K562/A02 cell line decreased progressively with increasing the concentration of PHI. Protein and gene level of MRP1 have no statistical significance (P&gt;0.05) no matter after or before PHI was used on different concentration.
Conlusion: The depletion effect of PHI on the intracellular GSH can not only partly enhance the reverse effect of ADM, but also enhance the sensitivity of K562/A02 cell line to ADM. Such depletion may diminish side effect and treatment dosage of ADM. It provides a new view to the therapy of leukaemia.
Treatment of Lower Risk MDS with Del 5q by Lenalidomide, with or without G-CSF: Current Results of the French Patient Named Program (ATU)
Authors: Lebras Fabien , Sebert Marie , Eclache Virginie , Lamy Thierry , Delaunay Jacques , Visanica Sorin , Dreyfus Francois , Banos Anne , Blanc Michel , Besson Caroline , Thomas Xavier , Rodon Philippe , Vey Norbert , Gardembas Martine , Bauduer Frederic , Ramee Jean-Francois , Delmer Alain , Rea Delphine , Turlure Pascal , Beyne-Rauzy Odile , Guerci Agnes , Ceccaldi Joel , Dugay J. , Fenaux Pierre , Ades Lionel 
Published: April 2020

Abstract
Background: LEN is very effective for the anemia of lower risk MDS with del 5q. A patient named program was launched by the French health agency (AFSSAPS) to use LEN in low and int 1 risk MDS with del 5q and transfusion dependence (&gt; 2 RBC units/2 months).
Methods: Patients received 10 mg of LEN/day, 3 weeks on, 1 week off. Hematological response (IWG 2006) was assessed after 8, 16 and 32 weeks of treatment. G-CSF use was recommended in case of grade 3 neutropenia, in order to avoid dose reduction, especially during the first 16 weeks of treatment.
Results: 71 pts from 35 centers were evaluable for response as of Aug 1st, 2008. Median age was 75 [range 43–93], 68% females, median interval from diagnosis to LEN treatment 28.6 months (range 1–235). At inclusion, 32 pts had del 5q syndrome, 10 RA, 13 RARS, 2 RCMD-RS and 14 RAEB-1. IPSS was low in 43% and int-1 in 57%. Del 5q was isolated, with 1 additional and &gt; 1 additional abn in 76%, 15% and 9% pts respectively. 23 pts were untreated, while 48 had previously received EPO. Median transfusion requirement was 4 units/2 months. 50/71 (69%) pts achieved erythroid response (IWG 2006), of whom 48 (67%) achieved transfusion independence (TI). Median time to TI was 12 weeks (range 8 – 32). TI was achieved in 76%, 62%, 68%, 70% and 50% pts with IPSS low, IPSS int 1, isolated del 5q, del 5q+1abn, del5q + &gt;1abn, resp, and 70% and 65% of EPO pretreated and EPO naive pts (p=NS). Grade III-IV neutropenia and thrombocytopenia were seen in 86% and 26% pts resp, leading to transient discontinuation followed by reduction of LEN dosing in 55%pts. Of the 19 pts who received GCSF, 3 (16%) required transient LEN discontinuation due to neutropenia compared to 15 (28%) of the 52 who did not receive GCSF (p=0.3). Apart from neutropenia and thrombocytopenia, grade III–IV side effects were Lyell’s syndrome (n=1), deep venous thrombosis (n=3) Quincke’s oedema (n=1), No patient died from sepsis due to neutropenia or bleeding due to thrombocytopenia. With a still short median follow up on treatment (28 weeks), no relapse had occurred. 1 pt (RAEB 1 with 7% of blasts, isolated del 5q and IPSS INT-1 at inclusion who had achieved TI) progressed to AML at week 48.
Conclusion: Our results confirm on a very multicenter basis the efficacy of LEN on the anemia of lower risk MDS with del 5q,. The use of G-CSF during the early treatment phase may somewhat reduce the proportion of early LEN discontinuation due to neutropenia.
Development of a Multiplex PCR Assay for the Detection of Genomic Copy Number Changes in Myelodysplastic Syndromes
Authors: Stamatoullas Aspasia , Waultier Agathe , Jardin Fabrice , Callat Marie Paule , Parmentier Francoise , Penther Dominique , Tilly Herve , Bastard Christian 
Published: April 2020

Abstract
Background: Myelodysplastic syndrome (MDS) is a heterogeneous group of disorders characterized by ineffective hematopoiesis. Clonal cytogenetic aberrations are found in about 50% of primary MDS and 80% of secondary MDS. The International Prognostic Scoring System (IPSS) introduces cytogenetic abnormalities as an independent prognostic factor for both survival and progression to AML. However, conventional cytogenetics or FISH substitutes are not available for all myelodysplastic patients. In order to overcome this problem, we developed a multiplex PCR assay (quantitative multiplex PCR of short fluorescent fragments or QMPSF) measuring the copy number of genes located in regions involved in myelodysplastic syndromes on chromosome 7 and 8 and on the 5q, 12p, 17p and 20q regions.
Material and method: In order to select target region, high resolution CGH (Agilent, 44K) was performed in 12 patients (5 complex karyotypes, 2 isolated monosomy 7, 2 isolated trisomy 8, 1 isolated 17p deletion, 1 isolated 20q deletion and 1 normal karyotype) for which DNA extracted from CD34+ sorted cells was available. Twenty three primer pairs were selected on 6 target chromosomes (5, 7, 8, 12, 17 and 20) in regions either commonly deleted or involved in biallelic deletions. A QMPSF assay was developed and applied on the first 12 patients. This assay was then applied on a series of 60 patients for which DNA extracted from either CD34 + sorted cells or total bone marrow (BM) cells or peripheral mononuclear cells (PB) was available.
Results: As previously reported by several groups, the correlation between cytogenetics and CGH was very good although high resolution CGH was able to detect small cytogenetically not detected defects in most patients. For the 23 QMPSF targeted genes, the correlation between CGH log ratio and QMPSF ratio was excellent. Using QMPSF, genomic copy number changes were detected in 21/27 CD34+ samples, 16/20 BM samples and 15/20 PB samples. However, when different samples were available for the same patient (CD34, BM and/or PB) the correlation between samples was not perfect and mainly depends on the MDS subtype, the % of blasts and the presence of cytogenetics abnormalities.
Conclusion: These results demonstrate that a single multiplex PCR assay targeting 23 genomic regions located on the 6 chromosomes most frequently involved in MDS can be helpful for the prognostic evaluation of MDS patients for whom cytogenetic data are not available, and that DNA extracted from PB or total BM cells can be informative in a substantial number of cases.
Clonal Heterogeneity in the 5q- Syndrome: NPMc+ and p53 Expressing Progenitors Are Insensitive to Lenalidomide Treatment and Expand at Disease Progression
Authors: Jädersten Martin , Saft Leonie , Pellagatti Andrea , Göhring Gudrun , Fernández-Santamaria Cristina , Wainscoat James S , Boultwood Jacqueline , Porwit Anna , Schlegelberger Brigitte , Hellström-Lindberg Eva 
Published: April 2020

Abstract
Clonal heterogeneity has not been described in patients with myelodysplastic syndrome (MDS) of the 5q- syndrome subtype, for which lenalidomide has emerged as a highly potent treatment. Interestingly, transformation to acute myeloid leukemia (AML) occurs more frequently in patients without a cytogenetic response to lenalidomide. We describe two patients with classical 5q- syndrome with complete erythroid and partial cytogenetic response to lenalidomide, who evolved to high-risk MDS and AML, respectively, with complex karyotypes including del(5q). Immunohistochemistry of pre-treatment marrow biopsies revealed small fractions of progenitors with aberrant cytoplasmic nucleophosmin (NPMc+) expression and in one patient also a TP53 mutation. Both lesions may induce a state of genomic instability, and represent novel findings in 5q- syndrome. These subclones remained stable during response, but expanded at transformation, suggesting that pre-existing cells with more malignant genotype may be insensitive to lenalidomide and responsible for disease progression. We are currently investigating a larger patient material for the presence of subclones with molecular lesions in order to assess the potential impact on the risk of leukemic evolution.
Characterization of Human Multiple Myeloma Cells with High-Resolution Magic Angle Spinning Magnetic Resonance Spectroscopy
Authors: Oriol Albert , Barba Ignasi , Barbera Angels , Arús Carles , Garcia-Dorado Jose-Luis , Ribera Josep-Maria 
Published: April 2020

Abstract
Advancements in the pathogenetic pathways in multiple myeloma have led to the identification of several primary and secondary genetic lesions and ultimately to a multiple myeloma genetic classification with prognostic implications. Although disregulation of cyclin activity has been recognized as a key event leading to the multiple myeloma phenotype, little is known about the metabolic consequences of this phenomenon. We have studied intact multiple myeloma cells by high resolution magnetic resonance spectroscopy to establish the metabolomic profiles of different native multiple myeloma cells as compared to other lymphoproliferative disorders.
Multiple myeloma cells obtained from bone marrow aspirates (n =15), blood (n =3) or other biologic tissues (n =2) from 20 multiple myeloma patients and separated by density gradient centrifugation were evaluated and metabolic profiles were correlated with cytogenetic characteristics of the disease and patients clinical characteristics.
Twelve patients were females (60%) with a median age of 65 years (range 50–82). Multiple myeloma monoclonal proteins were IgG (N=9), IgA (N=5) or BJ (N=6). Five of them (25%) had renal insufficiency. Nine patients (45%) had predominantly extramedullar diesase including four cases of plasma cell leukemia. IgH translocations were identified in 5 samples (25%), hyperploidy in 2 (10%), and other or no genetic lesions in 13 (65%), del13 was present in 9 samples (45%) and p53 alterations in 5 (25%).
Bone marrow samples from thirteen patients with conventional multiple myeloma presented a relatively constant metabolic pattern with predominantly lipidic signals and a metilen to metil ratio ranging from 1.9 to 4.9 (median 2.9). No differences in this pattern were observed among subgroups of primary translocations or involvement of Rb and p53 genes. Four patients with plasma cell leukemia and three with predominant extranodal disease presented either non detectable lipid signals (N=3) or a higher metilen to metil ratio ranging from 2.8 to 3.9 (median 3.5). In fact, extranodal or leukemic disease was significantly associated to undetectable lipids (P &lt; 0.031) or the composite variable undetectable lipids or metilen to metil ratio &gt; 3 (P &lt; 0.043). Furthermore, after a median follow-up of 18 months, absence of lipids in the metabolic profile was also associated to a shorter survival (median 0.45 years, 95%CI 0–1.03 versus 3 years, 95%CI 0.95–5.06, P &lt; 0.022).
These results suggest that metabolic profiles of different multiple myeloma genetic subtypes share common and reletively constant characteristics, while cells obtained from patients with plasma cell leukemia or predominantly extramedullar disease present a clearly distinct profile, probably reflecting the metabolic effect of clonal evolution at a genetic level.
The Prognostic Significance of 8p21 Deletion in Multiple Myeloma
Authors: Nahi Hareth , Sutlu Tolga , Alici Evren , Gahrton Goesta 
Published: April 2020

Abstract
In this study, we have investigated the prognostic impact of fluorescence in situ hybridisation (FISH) results for the chromosomal regions 1q21, 6q21, 8p21, 9p21, 13q14, 15q22, 17p13, 19q13 as well as the translocations t(4;14) and t(11;14) in patients with MM receiving ASCT. Randomly selected 65 patients were analyzed by FISH for these regions and hierarchical clustering analysis revealed associations between the occurrences of various chromosomal abnormalities. 58 patients received high-dose therapy (HDT) followed by autologous stem cell transplantation (ASCT) and 14 of these patients had monoallelic loss of the 8p21 region. Overall survivals (OS) and progression-free survivals (PFS) were significantly lower in patients with del(8p21). Moreover, multivariate analysis revealed that the occurrence del(8p21) is an independent prognostic factor for both OS and PFS. These findings suggest that the chromosomal region 8p21 could be monitored and used for the prognosis of patients with MM undergoing ASCT.
Alkylator-Induced DNA Damage Response in Multiple Myeloma (MM) Cells Is Amplified by Histone Deacetylase (HDAC) Inhibition, Resulting in Significant Synergism in Cell Death
Authors: Lee Choon Kee , Wang Shuiliang , Huang Xiaoping , Ryder John , Ordentlich Peter , Liu Bolin 
Published: April 2020

Abstract
One of the main mechanisms of action of HDAC inhibitors is the transcriptional reactivation of dormant tumor-suppressor genes through acetylation of histones, thereby inducing apoptosis. Treatment with HDACI has also been shown to induce chromatin destabilization in a transcription independent way. In the current study, we sought to determine whether HDAC inhibition induces DNA damage and amplifies alkylator-induced mitotic cell death in both melphalan sensitive- and resistant-MM cell lines (RPMI8226, 8226/LR5). The IC50 values of SNDX275, a class I HDACI agent, and melphalan on the 72-hour MTT assay were 268.05 nM and 245.94 nM in the RPMI8226, and 309.91 nM and 8657.46 nM in the 8226/LR5, respectively. When combined together at clinically attainable concentrations, the combination index by the Chou-Talalay method ranged from 0.27 to 0.75 for the RPMI8226 and from 0.33 to 0.7 for the 8226/LR5, indicating a powerful synergism. For elucidation of molecular mechanisms, MM1S and RPMI8226 cell lines were investigated for apoptosis, histone acetylation, cell cycle analysis, DNA double strand break and DNA damage response serially in 48-hour culture with SNDX-275 at 500 nM and melphalan at 10 μM, alone and in combination. Cleavage of PARP was seen following treatment with each SNDX275 and melphalan, but was highest at 48 hours with the combination of both. Apoptosis was associated with cleavage of caspases of 8, 3 and 9, which was most intense on combination. Melphalan amplified SNDX275-induced acetylation of H3. In cell cycle analysis by flow cytometry, SNDX275 caused an increase in G0-G1 and a decrease in S and G2-M. Cyclin D1, E2F-1 and p53 on western blot were not affected but expression of p21 increased. Melphalan arrested the cell cycle at G2, increased expression of p53 in the RPMI8226 and of p21 in the MM1S. The combination intensified the increase in p21 in both cell lines and in p53 only in the RPMI8226. Phosphorylation of H2AX, a marker of DNA double strand break, increased in a time dependent manner following each drug, along with an increase in phosphorylation of CHK1 and CHK2, indicative of initiation of DNA damage response. The increase in γH2AX and pCHK1 & 2, however, was considerably higher on combination than each drug alone. Furthermore, morphologic assessment of dead cells by the 48 hours of culture revealed a significant increase in mitotic catastrophe on combination in the MM1S: 0% on SNDX275 alone; 10% on melphalan alone; 43.4% on combination. The current study suggests that HDAC inhibition synergizes with melphalan in MM cells and that intensification of DNA damage is one of the mechanisms. Further studies are necessary to understand the role of HDAC inhibition for induction of mitotic catastrophe.
The Clinical Study of Combination of Bortezomib and Dexamethasone in Treatment of Newly Diagnosed Multiple Myeloma
Authors: Li Juan , Zeng Lijin 
Published: April 2020

Abstract
OBJECTIVE: To study retrospectively the response and side effects in two groups of patients with newly diagnosed multiple myeloma (MM) receiving bortezomib and dexamethasone regimen (VD) and vincristine combined with pirarubicin and dexamethasone and melphalan regimen (VADM).
METHODS: Nineteen patients were enrolled in a group of VD, receiving bortezomib 1.3mg/m2 on days 1,4,8,11 and dexamethasone 20mg on days 1–4 in a 21-day cycle. Blade Standard was used to evaluate the therapeutic effect and NCI-CTC was used to assess the adverse effect. Thirty-one matched patients with newly diagnosed MM who received VADM were used as a historical control group, receiving vincristine 0.4mg/d and pirarubicin 9mg•m−2•d−1 and dexamethasone 20mg/d and melphalan 12mg/d on days 1–4, with 28 days as a cycle.
RESULTS: During the median 9 months’ follow-up of patients who received VD, there were 73.7% patients(14/19)responding to the treatment, including 9 cases (47.4%) of complete remission or near complete remission. Light-chain type patients who received VD had a higher overall response rate and CR +nCR rate than that of the VADM (P&lt;0.05). The patients receiving VD who had renal inadequacy had an effective rate of 69.6% (5/6), which was similar to that of the others (69.2%, 9/13), and renal function relieve could be shown in the chemotherapy. The main adverse effects were fatigue, diarrhea, peripheral neuropathy, thrombocytopenia, and infection, with incidences of 73.8%, 68.4%, 63.2%, 31.5% and 26.3% respectively. Most of the adverse effects were mild and could be relieved by symptomatic treatments. The most common adverse event in the control group was neutropenia (83.8%), followed by infection(35.5%), vomiting (35.5%) and loss of hair(32.5%).
CONCLUSION: Although there was no significant difference of overall response rate between VD and VADM, VD has higher CR +nCR rate compared with VADM. VD can be tolerant in most patients, and is also safe in patients with renal inadequacy.
Screening JAK2 Exon 12 in JAK2 V617F Negative Patients with Myeloproliferative Disorders Reveals a New Splice Site Mutation
Authors: Bento C. , Estevinho A. , Rapado I. , Grande S. , Matos H. , Coucelo M. , Osorio S. , Albizua E. , Molinero R. , Duarte M. , Menezes C. , Kaeda J. , Martinez-Lopes J. , Ribeiro M. L. 
Published: April 2020

Abstract
Background. The identification of the somatic mutation V617F in exon 14 of the Janus kinase 2 gene (JAK2) has simplified the diagnosis of many patients affected with typical chronic myeloproliferative diseases (MPDs). In patients without the V617F the molecular basis of MPD are still unclear but, recently, mutations in exon 12 of the JAK2 gene have been identified in a minority of patients, associated with a selective increase in erythropoiesis resulting in polycythemia vera (PV) or idiopathic erythrocytosis (IE) (Scott et al, NEJM 2007).
Aim. To determine the JAK2 exon 12 mutational status in a group of JAK2 V617F negative patients with PV or IE.
Methods. Genomic DNA was extracted from peripheral blood leukocytes from 85 MPD patients (PV or IE) included in the present study. All the samples were tested for JAK2 V617F mutation by allele specific polymerase chain reaction (ASO-PCR) and those negative for V617F mutant allele were subjected to real time quantitative PCR (RQ-PCR) using hybridization probes. Subsequently, all JAK2 V617F negative samples by both ASO-PCR and RQ-PCR where subjected to direct sequencing to exclude JAK2 exon 12 mutations.
Results. JAK2 V617F was positive in 78 (91.7%) of the 85 patients, however, in two of these patients the V617F mutation was only detected upon subjecting genomic DNA to RQ-PCR which revealed low levels of the mutant allele, 2.65 % and 3.98%. Analysis of the JAK2 exon 12 in the seven JAK2 V617F negative patients detected three previously described mutations: a duplication (V536-1546) and two deletions (H539-K540del+542K and R541-E543 delinsK)and a previously unreported splice site mutation detected in intron 12 (IVS12nt6 T-C). To our knowledge this was the first description of intronic mutations in the JAK2 gene. No mutations were detected in the remaining 3 (3.6%) patients.
Conclusions. In this cohort study JAK2 mutations were observed in 82 (96,5%), of the 85 patients, of these 78 (95.12%) had the V617F allele. In two cases V617F was detected at low levels (2.65% and 3.98%) only by RQ-PCR, highlighting the need for sensitive techniques to detect somatic mutations. One of the patients, a young male with erythrocytosis and low serum Epo levels, revealed a previously unreported splicing mutation (IVS12 nt6: T-C). This study illustrates the heterogeneity at the DNA level in PV patients which may assist in better understanding the genotype and phenotype relationship in MPD patients and assist in further delineating the role of JAK2 in these disorders.
The Overall Analysis of JAK2 and MPL Mutation Status in Chinese MPD Patients by MassARRAY Assay
Authors: Zhang Sujiang , Li Jianyong , Shi Jingyi , Xia Jun 
Published: April 2020

Abstract
In recent three years the new notable progress associated with myeloproliferative disorder (MPD) was the identification of JAK2 and MPL mutations, which have been confirmed to be the major molecular mechanism and marker of BCR/ABL negative MPD. Multiple techniques including allele-specific PCR, conventional DNA sequencing, pyrosequencing as well as BsaXI restriction analysis have been used to detect these mutations. The previous data of Shanghai Group had demonstrated the sensitivity and preliminary result of JAK2 V617F in 162 Chinese patients with MPD using MassARRAY assay. To generally identify the frequency of these mutations in Chinese MPD patients, we continued to employ this technique-MassARRAY assay to detect JAK2 V617F, JAK2 K539L as well as MPL W515L mutation in a larger scale of Chinese MPD patients. A total of 204 Chinese MPD patients were enrolled in our study. These patients were referred to polycythemia vera (PV) (n=55), essential thrombocythemia (ET) (n=110), primary myelofibrosis (PMF) (n=29) and hypereosinophilic syndrome (HES) (n=10). 2 ml peripheral blood was obtained with informed consent and genomic DNA was isolated. The diagnosis of MPD was established according to the 2001 WHO diagnostic criteria. Finally, 187 patient samples with good signal can be analyzed for JAK2 V617F mutation. Among the available 101 signals of ET patients, 30 were heterozygous mutation and 9 were homozygous mutation. Among 52 PV patients, 25 were heterozygous mutation and 14 were homozygous mutation. Among 25 PMF patients, 9 were heterozygous mutation and 5 were homozygous mutation. None of HES patients was found harboring JAK2 V617F. In addition, we identified a PV patient harboring JAK2 K539L but not V617F mutation (1/52, 1.9%), and an ET patient harboring MPL W515L mutation (1/101, 1%). Both results were further confirmed by sequence analysis. Hence, we concluded that JAK2 V617F underlie the major molecular pathogenesis of Chinese MPD patients especially PV, however, JAK2 K539L and MPL W515L are rare events in these patients.