bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hematopoietic Microenvironment Regeneration Using Adipose-Derived Stem Cells.
Authors: Ogawa Rei , Fujimura Juri , Mizuno Hiroshi , Hyakusoku Hiko , Shimada Takashi 
Published: October 2019

Abstract
Backgrounds: We have studied hematopoietic microenvironment regeneration using adipose-derived stem cells (ASCs) harvested from mice. We showed that not only bone matrix but also bone marrow could be regenerated subcutaneously, in vivo. Transplanted ASCs proliferated and differentiated into osteoblasts in micropores on the surface of scaffolds made of hydroxyapatites (HA). We could regenerate a subcutaneous microenvironment in which hematopoietic cells survived, which may lead to new treatments for fibrotic bone marrow diseases, such as idiopathic myelofibrosis and osteopetrosis.
Objective: In this report, we examined the optimal shapes of scaffolds for bone and hematopoietic microenvironment regeneration using ASCs.
Methods: Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice, ASCs were isolated from the inguinal fat pads of GFP transgenic mice. ASCs were cultured in vitro and after three passages were seeded into the small pores of scaffolds of various shapes made of HA. The ASC-containing scaffolds were subsequently implanted into isogenic mice subcutaneously. Two months later, the scaffolds were extirpated for histological, immunohistochemical and flow cytometry analyses.
Results: Histological examination showed that the pores were filled with bone matrix and that the bone marrow was composed of adipocytes, hematopoietic cells, and vasculatures. Bone marrow regeneration on small, thin scaffolds of high porosity was better than on other scaffolds. Immunohistochemical analysis confirmed that the GFP+ ASCs that had differentiated into osteoblasts were composed of bone matrix. Flow cytometry analyses showed that the bone marrow was composed of blood cells, including populations of monocytes, lymphocytes and granulocytes.
Conclusions: Hematopoietic microenvironment engineering has great potential for hematopoietic disease therapy. The regeneration of the hematopoietic system using ASCs might be useful in the future for treating hematopoietic stem cell diseases such as leukemia as well as fibrotic bone marrow diseases.
Clinical and Experimental Study in 21 Cases of Acute Leukemia with Trisomy 4.
Authors: Pan Jin L. , Xue Yong Q. , Qiu Hui Y. , Zhu Yong J. , Wu Ya F. , Wang Yong , Shen Juan 
Published: October 2019

Abstract
To explore the clinical and experimental features of acute leukemia(AL) with trisomy 4, a retrospective study on the clinical and laboratory data in 21 cases of AL with trisomy 4 was performed. Chromosomes were prepared using direct method and/or short-term(24h) cultures of bone marrow cells. Karyotypic analysis was carried out using R- banding techique. 14 cases were studied by interphase FISH using a chromosome 4-specific α-satellite DNA probe labeled by spectrum Green to ascertain the presence of a clone with trisomy 4. 5 cases with t(8;21) revealed by karyotypic analysis were detected by dual-color FISH using t(8;21) translocaton probe to confirm the AML1/ETO rearrangement. The patients with AL and trisomy 4 had a median age of 50 years and a median survival of 10 months. They all had de novo AL expect two cases with secondary AL. M2 was the most frequent FAB subtype in this series (9/21 cases). The initial leukocyte count more than 10×109/L was seen in 16 cases. An enlargement of liver, spleen and/or lymph nodes in varying degrees was found in 15 cases. Among 15 cases received immunophenotypic analysis, 11 cases showed CD34 positivity and 6 cases co-expressed myeloid and lymphocytic antigens. Karyotypic analysis disclosed clonal trisomy 4 in 18 cases and one cell with +4 in 3 cases. Isolated trisomy 4 was found in 7 cases, while 14 cases had other abnormalities besides trisomy 4 including numerical abnormality (+6) in one case, and structural abnormalities in 13 cases among which t(8;21) was found in 8 cases. Interphase FISH confirmed that all 13 cases including 3 cases having one cell with +4 on karyotypic analysis had clonal trisomy 4. Dual-color FISH confirmed that all 5 cases with t(8;21) had AML1/ETO rearrangment. So we concluded that AL patients with trisomy 4 had unique clinical features and a poor prognosis.
Arsenic Trioxide Induces Mitochondrial DNA Mutation on D-Loop Region in Patients with Acute Myeloid Leukemia.
Authors: Zhou Jin , Meng Ran 
Published: October 2019

Abstract
Objective:To investigate the effect of arsenic trioxide (As2O3) on mitochondrial DNA (mtDNA) in acute myeloid leukemia cells.
Methods: NB4 cell line and primary leucocytes, isolated from peripheral blood of eight cases with newly diagnosed acute myeloid leukemia (the ratio of the blast cell was more than 80%), were incubated in RPMI-1640 medium supplemented with 10% heat-inactivated FCS, and treated with 0.1, 1.0, 2.0 micromol/L of As2O3 for 48hrs respectively in vitro. Blood samples in eight cases above were collected before and 14 days after As2O3 treatment with the dosage of 0.16mg/kg.d in vivo. Genome DNA was isolated from the cells and blood samples by salt fractionation, and the D-loop fragment of mtDNA was isolated from the genome DNA and amplified by polymerase chain reaction, and the sequencing techniques were applied to identify positive clones. MTT assay, electrophoresis of genomic DNA, and protein/DNA dual parameter flow cytometry were used to examine the effect of As2O3 on cell proliferation and apoptosis.
Results: mtDNA mutations were found on D-loop fragment of mtDNA in both NB4 cells and primary leucocytes, the number of single nucleotide polymorphism (SNP) or mutation were increased remarkably after 1.0, 2.0 micromol/L of As2O3 treatment both in vitro and in vivo than non-As2O3 treated baseline. The types of mutation were included the base transversion, transition, depletion and insertion. The 1.0 and 2.0 micromol/L of As2O3-treated samples revealed apoptosis, reduced proliferation and mtDNA mutation, while in the 0.1 micromol/L of As2O3-treated samples, showed reduced proliferation, differentiation and mtDNA mutation, but no apoptosis was appeared.
Conclusions: As2O3 promoted leucocytes’ mtDNA mutation on D-loop fragment both in NB4 cell line and in primary leucocytes isolated from patients with acute myeloid leukemia. Mitochondrial DNA might be one of the targets of As2O3 act on leukemia cells. Mitochondrial DNA mutation as well as nuclear DNA might participate the process of As2O3 inducing differentiation and reducing proliferation.
Clinical Relevance of Multidrug Resistance Proteins Expression in Patients with De Novo Acute Myeloid Leukaemia.
Authors: Podolak-Dawidziak Maria , Dus Danuta , Kielbinski Marek , Paprocka Maria , Haus Olga , Kuliszkiewicz-Janus Malgorzata , Kuliczkowski Kazimierz 
Published: October 2019

Abstract
Background. Early occurrence of the multidrug resistance (MDR) in de novo acute myeloid leukaemia (AML) may contribute to the treatment failure.The aim of this study was to investigate prospectively the clinical significance of the MDR proteins overexpression and their functional augmenting, in the context of other AML prognostic factors, such as age, immunophenotype and cytogenetic profile. We examined expression of MDR proteins: MDR1, MRP1, MDR3, BCRP, LRP and GSTπ and performed drug resistance functional assay in peripheral blood blasts of 25 patients with de novo AML at diagnosis and after the first chemotherapy cycle consisting of a 3 + 7 combination of DNR/Ara-C. MDR proteins presence and their functional activity (measured by fluorescent Rh123 dye efflux) were estimated by flow cytometry.
Results: Thirteen out of the 25 AML patients (52%) attained a CR with induction treatment, one had PR and eleven did not achieve remission. Out of 10 patients without CR, 2 died in aplasia and 9 were classified as an early death due to disease progression. All patients who achieved remission were younger than 55 years. Among 11 patients without remission, 9 expressed CD34; 6 of them had intermediate and 5 unfovorable cytogenetic profiles. In 10 out of 25 patients (40%) overexpression od MDR1 was shown at diagnosis, and was irreversible in 9 of them, those with poor clinical outcome (6 did not achieve CR, 1 had PR and one who obtained CR relapsed), whereas one patient who reversed achieved CR. The functional MDR assay showed the increased Rh123 efflux, both at diagnosis and after the first chemotherapy cycle in 12 patients (48%) and it influenced patients’ outcome in similar manner as MDR1 expression. At diagnosis other MDR proteins were also elevated in some AML patients:GSTπin 19 (76%), LRP in 9 (36%), MRP in 4 (16%) and MDR3 in 2 (8%). Seven AML patients (20%), both at diagnosis and after the first chemotherapy cycle co-expressed MDR1 with other multidrug resistance proteins and had enhanced Rh123 efflux. This co-expression resultes in 6 of them in chemotherapy resistance; the remaining one who achieved remission was young and had favorable cytogenetic profile. The results obtained demonstrate that the advanced age, unfavorable cytogenetic profile, overexpression of MDR1 at diagnosis and co-expression of other MDR proteins together with their functional activity contribute to the treatment failure in de novo AML.
Prevention of Homoharringtonine Induced Apoptosis by Antioxidants in K562 Cells.
Authors: Mai Wenyuan , Lin Maofang 
Published: October 2019

Abstract
Homoharringtonine (HHT) is a cephalotaxine ester derived from an evergreen tree found wildely throughout southern China, which has antileukemic activities against a variety of acute myeloid leukemic cells. HHT was recently shown by us to have differentiative and apoptotic effects on human leukemic cell lines. Recently, apoptosis has been associated with oxidative stress in a number of cell systems. Here we assessed the inhibitory capacity of antioxidant on HHT-induced apoptosis in the human leukemic cell line, K562. The apoptosis was analyzed by agarose gel electrophoresis of DNA, flow cytometry and terminal deoxyribonucleotidyl transferase-mediated dUTP-biotin nick labeling. K562 cells were treated with HHT at doses of 0ug/ml, 25ug/ml for 24 hours. Vitamin C (0.2 ug/ml) were added to medium 16 hour before addition of HHT. Results showed vitamin C inhibited oxidative stress and K562 cells apoptosis in HHT-treated groups (P < 0.05). Our data indicated that HHT treatment primarily induces K562 cells apoptosis at concentration up to 100 ug/ml. The underlying mechanism appears to involve mitochondrial damage via an increase in oxidative stress.
Downregulation of Survivin by Arsenic Trioxide in Acute Myeloid Leukemia Cell Line.
Authors: Silva Karina Lani , de Freitas Prazeres Martina , Maia Raquel Ciuvalschi 
Published: October 2019

Abstract
Caspases are proteins that play a central role in apoptosis. Therefore, triggering apoptosis through chemotherapeutical caspases inductor drugs is the major path in cancer treatment. However, hindering apoptosis by inhibitor of apoptosis proteins (IAPs) overexpression, have been described in many cancer types including leukemia and, is frequently related to drug resistance. Survivin, a member of IAPs family, is expressed in most human cancers but undetectable in the majority of normal adult tissues. In acute myeloid leukemia (AML), Survivin expression has been correlated with poor prognosis and chemotherapy resistance. These characteristics make Survivin eligible for a promising target for AML treatment. To explore the relationship between Survivin and drug resistance we investigated the alteration of Survivin expression in two AML cell lines HL60 (AML-M2) and U937 (AML-M5) and one chronic myeloid leukemia cell line in blast crisis for M6 (K562) treated with two chemotherapeutic drugs used in leukemia treatment: arsenic trioxide (As2O3) and doxorubicin (Dox). MTT assay was performed to determine the dose of drugs capable to induce cell death in 50% of treated cells (DL50). To verify the percentage of apoptosis induced by As2O3 and Dox at DL50 concentrations determined by MTT, the annexin V/propidium iodide-staining assay was performed and analyzed by flow cytometer. Western blot was used to analyze Survivin expression before and after drugs treatment at DL50 concentrations. Among the cell lines studied, HL60 was the most sensitive for both drugs tested. The DL50 concentrations obtained for As2O3 were 2, 4 and 5 μM at 24 hours for HL60, U937 and K562, respectively. Dox DL50 concentrations were 10 μM at 24 hours for HL60, 5 μM at 48 hours for K562 and 1 μM at 72 hours for U937. The annexin-V/PI staining showed that these drugs were capable to induce apoptosis in all cell lines tested. The percentages of apoptosis induction for As2O3 were 50% for HL60, 21,84% for U937 and 32,7% for K562 in comparison with control cells, while for Dox, the index of apoptosis were 86,8%, 35,7% and 2,2% for HL60, U937 and K562, respectively. Interestingly, at DL50 concentrations As2O3 and Dox inhibited Survivin expression in 72,7% and 69,2%, respectively. No significant alteration in Survivin expression was observed in U937 and K562 cell lines. Thus, HL60 cell line was the most sensible cell line studied and it was correlated with downregulation of Survivin expression. It suggests that Survivin could be considered a therapeutic target for AML-M2 and that As2O3 and Dox are capable to decrease drug resistance. However, the mechanism of action of As2O3 and Dox in Survivin expression seems to be cell type specific.
Trisomy 22 as the Sole Abnormality Is an Important Marker for Inversion 16 in Acute Myeloid Leukemia.
Authors: Li Jianyong , Zhou Huifen , Chen Lijuan , Pan Jinlan , Qiu Hairong , Xu Wei , Xue Yongquan , Lu Hua 
Published: October 2019

Abstract
Inv(16) has been reported in 10%~12% of acute myeloid leukemia (AML), mostly being associated with the M4Eo subtype, and is associated with a relatively favorable outcome. However, it is a cryptic rearrangement and often difficult to recognize in conventional cytogenetics (CC). Trisomy 22 is an uncommon karyotypic aberration in AML and is often associated with inv(16)(p13q22). In order to explore the value of trisomy 22 in the diagnosis of AML with inv(16), dual-color interphase fluorescence in situ hybridization (FISH) was performed in 19 AML cases with trisomy 22 abnormality. The probe was two-color break apart probe for CBFb with SpectrumRed on the centromeric side and SpectrumGreen on the telomeric side. And the results were compared with that of R-banding CC. CC did not reveal inv(16) in any of the 19 AML with trisomy 22, but FISH analysis showed inv(16) in 11 cases and del(16)(q22) in one case. Among 11 cases with inv(16), 9 were trisomy 22 as the sole abnormality, one was complicated with trisomy 8, and one was del(16)(q22). Four AML patients with trisomy 22 and inv(16) were analyzed by multiplex FISH (M-FISH) which revealed trisomy 22 only. This study further confirmed that trisomy 22 as the sole abnormality can be regarded as an important marker for the inv(16) in AML.
Fluorescence In Situ Hybridization Studies in 22 Cases of Plasma Cell Leukemia.
Authors: Li Jianyong , Xu Wei , Chen Lijuan , Zhu Yu , Pan Jinlan , Qiu Hairong , Xue Yongquan , Lu Hua 
Published: October 2019

Abstract
Background Plasma cell leukemia (PCL) is a rare malignant plasma cell disorder that usually carries an aggressive course with a rapidly fatal outcome. Cytogenetic studies performed on plasma cell disorder are scarce and difficult because of the low proliferation rate of plasma cells. Fluorescence in situ hybridization (FISH) analysis is an attractive alternative for evaluation of numerical and structural chromosomal changes in PCL.
Methods Interphase FISH (I-FISH) and two different specific probes for the regions containing 13q14.3 (D13S319) or 14q32 (IGHC/IGHV) were used to detect 13q14 deletion [del(13q14)] or IgH rearrangement in 22 PCL patients. For patients with IgH rearrangement, probes for IgH(JH) and 11q13 (CCND1) or 4p16 (FGFR3) were used to detect t(11;14)(q13;q32) or t(4;14)(p16;q32).
Results Molecular cytogenetic aberrations were found in 19 of 22 (86.4%) PCL patients. Del(13q14) was detected in 13 cases (59.1%), and IgH rearrangement in 17 (77.3%) patients including 7 with t(11;14) and 3 with t(4;14). 14q32 rearrangement and 13q14 deletion were found concurrently in 11 cases (50%).
Conclusions Chromosomal abnormalities are frequent in PCL. The most frequent aberrations among the cases was the 14q32 rearrangement and 13q14 deletion. I-FISH technique is useful to detect molecular cytogenetic aberrations and should be used in the routine evaluation of PCL.
Profound Hyperbilirubinemia during Induction of Adult Acute Lymphoblastic Leukemia: Capping L-Asparaginase Dose to 10,000 Units Attenuates It without Compromising Remission Rate.
Authors: Vora Amish D. , Pai Vasnat R. , Kumar Prabhash , Banavali Shripad , Nair Reena , Kurkure Purna , Parikh Purvish M. 
Published: October 2019

Abstract
Introduction: Hyperbilirubinemia is a well known side-effect of L-asparaginase. Grade 4 hyperbilirubinemia (bilirubin > 3) results in delayed chemotherapy; compromising dose intensity. Hence a strategy to prevent this is important.
Materials & Methods: At our center, we see around 125 cases of newly diagnosed adult acute lymphoblastic leukemia (age>24 years) every year. Since 2005, we have started treating these adult patients on modified BFM 90 protocol. This protocol includes 8 doses of L-asparaginase at 10,000 units/m2 during induction. Hyperbilirubinemia of grade 4 (median bilirubin upto 20) was encountered in the first few patients. To avoid delay in chemotherapy, we modified the protocol to cap L-asparaginase dose at 10,000 u for all patients.
Results: A total of 64 patients with ALL was treated on this modified protocol in 18 months. We compared the degree of hyperbilirubinemia in this group of patients with the first 20 patients who were treated with the standard dose of L-asparaginase. Median age of the full cohort (All 84 patients) was 34 years (24–60). Fifty four patients had precursor B-cell ALL while 30 patients had T-cell ALL. Male: Female ratio was 4.1:1. All the analysed patients have completed induction. In first group of 20 patients who had received standard dose of L-asparaginase, 6 patients developed grade 4 hyperbilirubinemia (median bilirubin 23). SGPT, SGOT and SAP were either normal or borderline elevated. Viral markers at diagnosis and at the onset of hyperbilirubinemia were negative. No significant hepatic abnormality was seen on USG abdomen. In four out of 6 patients’ bilirubin returned to normal after conservative management, over a period of 3 weeks. The two remaining patients developed sepsis, had deranged coagulation parameters and succumbed to sepsis. Of the second group of 64 patients ( L-asparaginase at 10,000 fixed dose), 5 patients developed hyperbilirubinemia (highest value of bilirubin 4). All other investigations (listed above) were normal.All patients’ bilirubin came back to normal within 1 week of interruption of chemotherapy. Complete remission rates were 90% in the first group of 20 patients and 92% in the second group of 64 patients.
Conclusion: In the cohort of 64 patients (second group), capping of L-asparaginase resulted in significant reduction in intensity, incidence and duration of hyperbilirubinemia. This modification did not affect CR rate, though we are currently evaluating long term survival to confirm sustained efficacy.
Remission Induction with Cladribine, Cytarabine and Idarubicin (CAI) in Elderly Patients with Relapsed AML - A Pilot Study.
Authors: von Lilienfeld-Toal Marie , Hahn-Ast Corinna , Orlopp Katjana , Ingo Schmidt-Wolf G.H. , Axel Glasmacher 
Published: October 2019

Abstract
Elderly (>55 years) patients with relapsed acute myeloid leukemia (AML) generally have a poor prognosis mainly due to a low rate of 2nd complete remission (CR). Commonly, salvage therapy consists of regimens including fludarabine. However, results have been disappointing and the efficacy of fludarabine in AML is questionable. In contrast, there is evidence that 2CdA, another purine analogue, has single drug activity in AML as well as enhancing the effect of other cytostatic drugs such as ARA-C. Here we present data from a pilot study evaluating CAI: Cladribine (2CdA) in addition to cytarabine (ARA-C) with idarubicin (Ida) as a salvage therapy in relapsed AML.
Patients and Methods: Two treatment protocols were tested from May 2005 to May 2006 at our institution:2CdA 5 mg/m2/12 h, d 1–5, ARA-C 1000 mg/m2/12 h, d1-5 and Ida 12 mg/m2/d, d1-32CdA 5 mg/m2/12 h, d 1–3, ARA-C 1000 mg/m2/12 h, d1–3 and Ida 8 mg/m2/d, d1–3.Protocol A was designed for patients < 60 years and considered fit for intensive therapy.
Results: Nine patients were included in the study after written informed consent. Eight of them are evaluable to date. Three patients received protocol A (median age 57 years), whereas 5 received protocol B (median age 65 years). Although one patient died during neutropenia and one patient had refractory disease, response after one cycle of CAI was favourable in 6 patients: 4 achieved CR and 2 CR with incomplete regeneration (response rate 75%). Of those, 3 patients went on to receive further consolidation therapy with a second cycle of CAI, one with ARA-C/Ida and one with 2CdA/ARA-C. Toxicity was predominantly neutropenia with subsequent infectious complications: median duration of neutropenia was 28 days (IQR 22–34) with all patients experiencing febrile neutropenia (median duration 8 days IQR 3–9) and 4 episodes of septicemia. The second course of CAI resulted in an even longer duration of neutropenia: 37 days (IQR 16–50) with all patients having septicaemia. Of the 6 responding patients 2 died subsequently, one of a second relapse and the other of neutropenic sepsis after the 2nd course of CAI. The other 4 patients are still alive (follow-up 4 months to 1 year).
Next steps: Because of these promising results we decided to pursue the study but change the protocol as follows:
Patients will only be treated according to protocol B, application of growth factors from day 15 onwards will be mandatory, antibiotic prophylaxis with a chinolone and twice weekly Cotrimoxazole will be mandatory and a second cycle will omit idarubicin.
Conclusion: Combination therapy with cladribine, cytarabine and idarubicin appears to be feasible and successful in elderly patients with relapsed AML. However, infections are a serious complication warranting intensive supportive care.