Objective: To study cytotoxicity and antineoplastic effect in vitro Solanum nigrum L extract on U266. Methods: U266 cells were cultured together with the extract of Solanum nigrum L. Cytotoxicity assay was tested by CCK-8. Cell cycle and apoptosis were determined using flow cytometry (FCM) analysis. Results: Extract of Solanum nigrum L showed strong cytotoxicity against U266 cells. The IC_ 50 was about 117 mg/L. After exposure of U266 cells to the drug for 48 hours, the cell cycle distribution was changed compared with the controls. There was decrease of cells in the G_0/G_1 phase with increase of cells in the S phase and G_2/M phase. Apoptosis of U266 cells could be shown with staining of Annexin V FITC/PI or TFAR19 testing through FCM. The proportion of apoptotic cells increased in parallel with the increase of the drug dosage. Conclusion: Solanum nigrum L extract showed strong cytotoxicity effect on U266 cells. The antineoplastic effect of the drug can partly be ascribed to its apoptotic inducing effect.

Objective To enhance engraftment by infusing two partially matched cord blood (CB) units from unrelated donors simultaneously. Methods Two patients with high-risk leukemia(1 case of ALL and 1 case of CML-BC) received total body irradiation with antithymocyte globulin (ATG) and then were transplanted with two HLA-mismatched CB units from unrelated donors. A combination of cyclosporine A, methylprednisonelone and mycophenolate mofetil (MMF) was administered for graft-versus-host disease (GVHD) prophylaxis. Results Two patients were all engrafted and the time to neutrophil≥ 0.5 ?10 9/L and platelet≥20?10 9/L were 21, 22 days and 51, 28 days post-transplant respectively, and DNA analysis showed only one donor DNA was detectable after engraftment. There was no serious GVHD. And these 2 patients are still alive and disease free for more than 34 and 30 months repectively. Conclusion Double units cord blood from unrelated donors can restore hematopoiesis and is suitable in adults without bone marrow donors.

Objective: To explore the effect of simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (the rate-limiting enzyme of cholesterol synthesis) on the proliferation and apoptosis of chronic myelogenous leukemia (CML) cells. Methods: Both normal and CML bone marrow progenitor cells were assayed in semisolid methylcellulose culture after incubation for 24 hours in suspension culture with 10 mg*L－1 simvastatin. Also, sub-G1 cells and DNA end-labeling positive cells as apoptotic cells were identified by flow cytometry after being exposed to simvastatin for 72 hours. Results: Simvastatin selectively inhibited proliferation and induced apoptosis of CML cells, but had no much influence on normal bone marrow cells. Conclusion: CML cells are more sensitive to a short-term exposure to simvastatin than normal bone marrow cells. It will be a promisingly effective chemotherapeutic agent or in vitro purging agent in autologous stem cell transplantation for the treatment of CML.

Objective Detection of single nucleutide polymorphisms in CD31-563 codon by denaturing high-performance liquid chromatography(DHPLC). Methods The eighth exon fragments of CD31 code gene in chromosome 17q23 were amplified by PCR. The fragments were analysed with DHPLC and were sequenced and compared with the sequences available in National Center for Biotechnology Information ( NCBI) database. Results The length of the amplified fragments is 203bp. There are three kinds of pictures when the fragments are analysed by DHPLC, after comparing with direct sequencing, the pictures of G/G homozygous、A/A homozygous、G/A heterozygous are obviously different, thus the individuals with the three kinds of genetype can be distinguished accurately. In 74 healthy people studied,the gene frequencies of CD31-563S(AGC) is 0.514, the gene frequencies of CD31-563N( AAC) is 0. 486. Conclusion DHPLC can effectively, economically and accurately detect the single nucleutide polymorphisms in CD31 -563 codon.

Objective To evaluate the value of plastic-embedded bone marrow (BM ) biopsy sections in diagnosing myelodysplas-tic syndrome(MDS) .Methods HGF and Gomori stained .Glycol methacrylate(GMA) embedded BM biopsy specimens were detec-ted from 125 MDS patients with 25 cases in good health as control .Dyshematopoiesis and abnormal stroma cell responses were se-lected as main observed projects .Results Bone marrow biopsy sections gave an exact assessment of cellularity ,19 cases (15 .2% ) showed a decrease of cellularity as hypocellular MDS .66 cases(52 .8% ) of these cases did show abnormal localization of erythro-blastic clusters ,biopsy of 64(62% ) of low risk group 103 cases did show abnormal localization of immature precursors such as blasts and promyelocytes(ALIP positive) .Numeration of megakaryocytes and mast cells within 1mm2 acre of BM tissue were all much in MDS group than in control group .Gomori′s stain revealed moderate increase in reticulin fibres in 65 cases ,10 cases(8 .0% ) showed degree of myelofibrosis were + + + as hyperfibrotic MDS .Among 125 cases ,33 cases(26 .4% ) of bone marrow biopsies showed different types of erythrocytic and granulocytic immature cells with unusual intrasinusoidal bone marrow infiltration .Con-clusion It is clear that GMA embedded BM biopsy section is helpful in making diagnosis of MDS ,and has important clinical appli-cation value .

Objective To analyze the clinical and pathologic features in patients with ldiopathic cytopenia of undetermined signif-icance (ICUS) ,and provide the diagnostic basis .Methods 10 ICUS patients who did not fulfil minimal diagnostic criteria for MDS but suffered from constant (>6 months) progressive cytopenia ,with no marked dysplasia (<10% in three-major cell lineages) and abnormal karyotype were reported .Results 2 of 10 patients transited to MDS ,bone marrow review showed that ≥2 cells appeared diagnostic pathology development .Karyotype analysis showed that 1 case was still normal ,1 case lost a normal chromosome 20 (20q-) ,the rest 8 cases had been efollowed up for 6 to 25 months and still maintain ICUS .Conclusion Patients with MDS who fail to meet the diagnostic criteria of ICUS ,must carry out regular follow-up .There are some patients can transition to public MDS after a certain incubation period ,in fact it is Pre-MDS or MDS-early before ,which shows that ICUS should cause clinical attention ,so that avoid missed diagnosis .

The new wave of artificial intelligence pushed by deep learning algorithms has dramatically promoted the development of big data analysis technology. On the other hand, advances in life sciences represented by high-throughput genome sequencing have provided massive medical data. Artificial intelligence technology has also provided a powerful tool for hematological malignancy research. This article introduces related research progress in the 61st American Society of Hematology Annual Meeting.

Objective To study the incidence, risk factors and prognosis of central nervous system (CNS) complications after hematopoietic stem cell transplantation ( HSCT) in order to prevent or reduce its occurrence, provide better diagnosis and treatment and improve the survival of the patients.Methods A total of 640 patients who consecutively underwent HSCT in our hospital between May 2001 and December 2007 were included.The clinical outcomes of the patients who developed CNS complications were analyzed.Results The patients received stem cells from haploidentical family members ( Haplo, n = 289 ) , identical siblings (IS, n = 237) , unrelated donors ( URD, n = 83) , unrelated cord blood (n = 14) , syngeneic siblings (n = 9 ) or autologous peripheral blood ( n = 8 ).Fifty-seven of 640 patients (8.9% ) developed CNS complications.The incidences were 12.0%, 13.5% and 3.4% in URD-HSCT, Haplo-HSCT and IS-HSCT respectively ( P <0.001).The incidences of CNS complications were 19.4% and 8.3% in cases who received or did not receive conditioning with TBI ( P = 0.047 ).There was no significant difference in the incidences of CNS complications between children (15.3% ) and adults(8.3% ) (P = 0.072).Similar incidences of CNS complications were seen in patients with hematological malignancies (8.9%) and non-malignant hematological disorders (7.7%)(P = 1.000).Five of the 57 patients developed two kinds of CNS complications.The patterns of CNS complications included relapse (17 cases) , infections (15 cases) , cyclosporine or FK506 encephalopathy (9 cases) , cerebral hemorrhage ( 8 cases) , cerebral infarction (2 cases), Wernicke's encephalopathy (1 case), skull fracture (1 case), drug-related meningitis (1 case), hepatic encephalopathy (3 cases), post-transplant lymphoproliferative disorder (1 case) and undetermined causes (4 cases).The overall mortality in the patients who developed CNS complications was 57.9% and 66.7% of them died of CNS complications.Conclusions CNS complications are not uncommon after HSCT and they have high mortality and poor prognosis.Our data suggest that haplo-HSCT,URD-HSCT and conditioning with TBI, but not the age and types of hematological diseases are the risk factors for development of CNS complications.Relapse and infections are the most common CNS complications in HSCT recipients.Early diagnosis and appropriate management are crucial to the improvement of clinical outcomes in these patients.

OBJECTIVETo observe the conditioning regimen, efficacy and side effects of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for hemophagocytic lymphohistiocytosis (HLH).

METHODFrom 2010 to 2012, a total of 11 cases after allo-HSCT were evaluated including 8 cases with familial hemophagocytic lymphohistiocytosis (FHL) and 3 cases with Epstein-Barr virus (EBV) related HLH. Allo-HSCT from HLA haploidentical HSCT was performed for 3 cases and unrelated allo-HSCT for 8 cases; 7 cases underwent allo-HSCT with conditioning regimen of etoposide (VP16), busulphan (Bu), fludarabine (Flu) and antilymphocyte globulin (ATG) and 4 cases with Flu, melphalan (Mel) and ATG. Cyclosporine (CsA) or tacrolimus, mycophenolate (MMF) and methorexate (MTX) were used for prevention of graft versus host disease (GVHD). Four cases received anti-CD25 MoAbs, 7 cases received cord blood and 1 of them received haploidentical bone marrow to prevent GVHD.

RESULTThree cases died after allo-HSCT. The median overall survival time of the 8 cases evaluated was 585 days (154-1 115 d). All the patients were successfully engrafted. Acute GVHD (aGVHD) occurred in 8 cases, including 3 cases of gradeI/II and 5 cases of grade III/IV. Chronic GVHD (cGVHD) occurred in 4 cases. Seven cases had cytomegalovirus (CMV) reactivation.

CONCLUSIONThe allo-HSCT was successful in treating primary and refractory hemophagocytic syndrome.

Adolescent ; Child ; Child, Preschool ; Cyclosporine ; administration & dosage ; Cytomegalovirus Infections ; epidemiology ; prevention & control ; Female ; Graft vs Host Disease ; epidemiology ; prevention & control ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunosuppressive Agents ; administration & dosage ; Lymphohistiocytosis, Hemophagocytic ; mortality ; therapy ; Male ; Survival Rate ; Tissue Donors ; Transplantation Conditioning ; methods ; Treatment Outcome

Objective: To analyze the incidence and mutation characteristics of FLT3 gene mutation and clinical efficacy of tyrosine kinase inhibitor (TKI) in patients with mixed phenotype acute leukemia (MPAL). Methods: A total of 48 patients with MPAL who were admitted to Hebei Yanda Lu Daopei Hospital from June 2015 to February 2018 were retrospectively analyzed. The common mutated 58 genes in hematologic malignancies were detected by using amplicon-targeted next generation sequencing, of which internal tandem duplication (ITD) and point mutation occurred in the hotspot region of exon 14, 15 and 20 in FLT3 gene. Multiplex polymerase chain reaction (PCR) analysis was used to detect 35 gene fusions in hematological neoplams. Results: There were 7 cases of FLT3 mutation in 48 MPAL patients, which were all ITD mutations. The median length of the inserts of FLT3-ITD was 48 bp, and one MPAL patient carried 2 multiple length inserts simultaneously, and the median variant allele frequency (VAF) was 40.5% (7.9%-84.7%). There were no statistically significant differences in clinical and genetic characteristics between FLT3 mutation-positive and FLT3 mutation-negative MPAL patients (both P > 0.05). Among 7 FLT3 mutation-positive MPAL patients, 4 cases were often accompanied with RUNX1 mutation. A total of 4 MPAL patients with FLT3-ITD-positive received sorafenib or sunitinib combined chemotherapy, and 3 of them achieved complete remission. Conclusions ITD mutation is the main part in the FLT3 mutation of MPAL patients. FLT3-ITD-positive MPAL patients are often accompanied with RUNX1 mutation, which may benefit from targeted therapy with FLT3 kinase inhibitor.