Female ; Humans ; Leukemia, Myeloid, Acute ; complications ; etiology ; Middle Aged ; Myelodysplastic Syndromes ; complications ; etiology ; Uterine Cervical Neoplasms ; complications ; drug therapy ; radiotherapy

OBJECTIVETo study the effects of arsenic trioxide on apoptosis of peripheral T-lymphocytes from asthmatic patients and normal subjects in vitro.

METHODSThe T-lymphocytes were isolated from the blood of 21 asthmatic patients and 20 healthy controls and treated with arsenic trioxide and dexamethasone. Cell apoptosis was observed by fluorescence microscope and measured with flow cytometry and Cytochrome C ELISA kit.

RESULTSThe T-lymphocytes from the asthmatic patients, when compared to those from of the healthy control, exhibited decelerated spontaneous apoptosis after a 24-hour incubation in vitro. Dexamethasone treatment significantly increased the percentage of apoptotic T-lymphocytes from both asthmatic patients and normal subjects in comparable magnitude. Arsenic trioxide treatment, in contrast, significantly increased the percentage of apoptotic T-lymphocytes from asthmatic patients, but slightly affected the cells from the control group.

CONCLUSIONSSpontaneous apoptosis of T-lymphocytes can be decelerated in asthmatic patients, whose T-lymphocytes are more sensitive to arsenic trioxide-induced apoptosis than those of normal subjects, but the T-lymphocytes from normal subjects and asthmatic patients are equally sensitive to dexamethasone.

Adult ; Anti-Asthmatic Agents ; pharmacology ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Asthma ; blood ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Male ; Microscopy, Fluorescence ; Oxides ; pharmacology ; T-Lymphocytes ; drug effects ; pathology

OBJECTIVESTo evaluate the efficacy and safety of imatinib mesylate (imatinib) for myeloproliferative neoplasm (MPN) patients with eosinophilia.

METHODSEight MPN patients with eosinophilia and positive FIP1L1-PDGFR alpha gene and one CEL, NOS were treated with 100 mg or 400 mg/d imatinib orally.

RESULTSHematological remission (HR) rate was 100%, including complete HR (CHR) rate 87.5%, and partial remission (PR) rate 12.5% with a median follow-up of 16 (6.0 -26.0 ) months. Complete molecular remission (cMR) rate was 85.7%. The median time of FIP1L1-PDGFR alpha fusion gene becoming negative was 4 (1.5 - 8) months. Three patients withdrew imatinib after getting cMR. After a median follow-up of 12 months, all the 3 patients remained in CHR. The main adverse effect of imatinib was mild myelosuppression occurring in 37.5% of patients in a median time of 6 (4 - 9) days after treatment.

CONCLUSIONImatinib in treatment of MPN with eosinophilia and positive FIP1L1-PDGFR alpha gene patients can induce high hematologic and molecular remission. The adverse effects of imatinib are mild and tolerable.

Adult ; Aged ; Benzamides ; Eosinophilia ; complications ; Follow-Up Studies ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Myeloproliferative Disorders ; complications ; drug therapy ; genetics ; Oncogene Proteins, Fusion ; genetics ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; Treatment Outcome ; Young Adult ; mRNA Cleavage and Polyadenylation Factors ; genetics

Objective:This study aimed to investigate the effect of differentiation osteogenic bone marrow mesenchymal stem cells (De-BMSCs) transplantation on the promotion of bone formation at the tendon-bone interface after anterior cruciate ligament reconstruction (ACLR), and further explored the molecular mechanism of the enhanced osteogenic effect of De-BMSCs.Methods:BMSCs from femur and tibia of New Zealand White rabbit were subjected to osteogenic induction and then cultured in no osteogenic factor medium; the obtained cell population was termed De-BMSCs. De-BMSCs were induced into osteo-, chondro-and adipo-differentiation in vitro to examine the characteristics of primitive stem cells. ACLR model with a semitendinosus tendon were performed in 48 adult rabbits, three groups were established: control group with alginate gel injectionat the tendon-bone interface, BMSCs group with the injection of alginate gel containing BMSCs, De-BMSCs group with the injection of alginate gel containing De-BMSCs. At 4 and 12 weeks after surgery, rabbits in each group were sacrificed to evaluate tendon-bone healing by histologic staining, micro-CT examination, and biomechanical test. During osteogenic differentiation of De-BMSCs, si-RNA of nuclear factor of activated T cells 2 (NFATc2) si-RNA of nuclear factor of activated T cells 1 (NFATc1) were used to verify the molecular mechanism of enhanced osteogenic effect of De-BMSCs.Results:De-BMSCs exhibited some properties similar to BMSCs including multiple differentiation potential and cell surface marker. At 4 weeks after surgery, the BV/TV value of the De-BMSCs group 0.36±0.01 was significantly higher than that of the control group 0.24±0.03 and BMSCs group 0.30±0.02 (all P<0.05), and the maximum load 40.34±1.19 N and stiffness 20.67±2.14 N/mm were significantly higher than those in the control group 14.88±2.74N, 8.67±2.19 N/mm and the BMSCs group 26.31±1.76 N, 13.81±2.14 N/mm (all P<0.05). At 12 weeks after surgery, the BV/TV value of the De-BMSCs transplantation group 0.47±0.02 was significantly higher than that of the control group 0.30±0.02 and the BMSCs group 0.35±0.03 (all P<0.05), and the maximum load 64.46±6.69 N and stiffness 25.18±3.11 N/mm were significantly higher than those in the control group 41.01±6.12 N, 11.59±2.54 N/mm and the BMSCs group 48.21±4.12 N, 15.89±2.94 N/mm (all P<0.05). During the osteogenic differentiation of De-BMSCs, the expressions of Nanog and NFATc1 were synergistically increased which promoted interaction of NFATc1 and Osterix ( P< 0.05), resulting in the increased expression of osteoblast marker genessuch as COL1A, OCN, OPN (all P< 0.05). Conclusion:De-BMSCs transplantation could promote bone formation at the tendon-bone interface after ACLR,Nanog/NFATc1/Osterix signaling pathway mediated the enhancement of the osteogenic differentiation effect of De-BMSCs.

OBJECTIVETo investigate JAK2 exon 12 mutations in patients with Philadelphia (Ph) chromosome-negative myeloproliferative neoplasms (MPN) and the clinical characteristics of patients with JAK2 exon 12 mutants.

METHODSAllele-specific PCR (AS-PCR) was applied to identify JAK2 V617F mutation. Genomic DNA corresponding to exon 12 of JAK2 gene and epigenetic regulator gene (TET2, ASXL1, EZH2) were amplified by polymerase chain reaction (PCR). Identification of mutants was by direct sequencing and classification of mutation types by sequencing followed by plasmid cloning. SNP genotyping of two 46/1 tag SNPs, rs12340895 and rs10974944, was analyzed using commercially available Taqman assays on the 7500HT real-time PCR instrument according to standard protocols.

RESULTSNo JAK2 exon 12 mutation was detected in patients with ET, PMF or JAK2 V617F positive PV. Among 13 JAK2 V617F negative PV patients, JAK2 exon 12 mutation was detected as N542-E543del in 2(15.4%) patients who presented with a phenotype of predominant erythrocytosis and erythroid colonic grown from their bone marrow samples in the absence of exogenous EPO, reduced serum erythropoietin (EPO) level, and no mutations in TET2, ASXL1 or EZH2 genes. One of the affected patients was heterozygous for 46/1 but the second was negative for this haplotype.

CONCLUSIONThere was no need to detect JAK2 exon 12 mutation in ET, PMF or MPN-U patients without JAK2 V67F mutation. Ph negative MPN patients with JAK2 exon 12 mutations had somewhat unique clinical and laboratory features.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow Neoplasms ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Myeloproliferative Disorders ; genetics ; Philadelphia Chromosome ; Young Adult

BACKGROUNDHepatic fibrosis is the key stage of the pathological progress from hepatic injury to cirrhosis. Ursodeoxycholic acid (UDCA) has been known as having significant clinical therapeutic effects on chronic liver diseases. Our research aimed to study the effect of UDCA on the signaling pathway of transforming growth factor beta1 (TGFbeta1)/Smad and discuss its possible molecular mechanisms of inhibiting hepatic fibrosis.

METHODSRat hepatic stellate cells were cultured in vitro and randomly assigned to 4 groups. Group A was control group, with only DMEM culture medium applied, and groups B, C, D were experimental groups, with different doses of UDCA (1.0 mmol/L, 0.5 mmol/L and 0.25 mmol/L respectively) added into their DMEM culture medium for further culture of 24 hours and 48 hours. The protein expressions of TGFbeta1, TGF type I receptor, Smad3, Smad4 and Smad7 were measured by Western blotting, as well as the expressions of TGFbeta1, Smad3, Smad7 and cAMP response element (CREB) binding protein (CBP) mRNA by real-time PCR. SPSS 11.5 statistical package was adopted for data analyses.

RESULTSCompared with control group, the mRNA expressions of TGFbeta1 in the high and middle UDCA dose groups for 24 hours and 48 hours significantly decreased (P < 0.05), the protein expressions of TGFbeta1 in the two above groups for 48 hours and in the high dose group for 24 hours significantly decreased (P < 0.05). The protein and mRNA expressions of Smad3 in each UDCA dose group for 24 hours and 48 hours significantly decreased, with significant difference among different UDCA dose groups and between that of 24 hours and 48 hours observed (P < 0.05). The protein and mRNA expressions of Smad7 in the high and middle UDCA dose groups for 24 hours and 48 hours significantly increased. The CBP mRNA expression in each UDCA dose group for 24 hours and 48 hours significantly decreased (P < 0.05), with significant difference among different UDCA dose groups observed (P < 0.05).

CONCLUSIONUDCA could curb the development of hepatic fibrosis through affecting the signaling pathway of TGFbeta1/Smad by inhibiting the expressions of TGFbeta1, Smad3 and CBP and increasing the expression of Smad7.

Animals ; Blotting, Western ; Cells, Cultured ; Cholagogues and Choleretics ; pharmacology ; Cyclic AMP Response Element-Binding Protein ; genetics ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Polymerase Chain Reaction ; Rats ; Receptors, Transforming Growth Factor beta ; metabolism ; Signal Transduction ; drug effects ; Smad Proteins ; metabolism ; Smad3 Protein ; genetics ; metabolism ; Smad4 Protein ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Ursodeoxycholic Acid ; pharmacology

OBJECTIVETo investigate the relationship between DNA homologous recombination (HR) repair genes RAD51-G135C/XRCC3-C241T polymorphisms and development of acute myeloid leukemia (AML) with recurrent chromosome translocation.

METHODSGenomic DNA was extracted from bone marrow cells of 625 de novo AML patients and peripheral blood cells of 806 patient family members and 704 unrelated volunteers. Genotypes of RAD51-G135C and XRCC3-C241T were analyzed by PCR-RFLP. Cell lines with genotypes differed from XRCC3-C241T were selected and irradiated in vitro. The CBFβ-MYH11 fusion gene was detected by TaqMan real-time PCR.

RESULTSThe XRCC3-C241T variant (C/T + T/T) showed 6.22-fold and 6.99-fold increase in the risk of developing the AML with inv(16)/t(16;16)/CBFβ-MYH11 as compared with the volunteer and family member controls respectively; the RAD51-G135C homozygote-type (C/C) variant showed 0.87-fold (P = 0.010) and 1.15-fold (P = 0.001) respectively increase in the risk of this subtype AML. In the irradiated group, the CBFβ-MYH11 mRNA level in HL-60 cells was 59.49 times increased than that in KG1a cells. However, the RAD51-G135C and XRCC3-C241T variants had no correlations with the risk of development of t(15;17)/PML-RARα(+)AML, t(8;21)/AML1-ETO(+) AML and 11q23 AML subtypes.

CONCLUSIONThe XRCC3-C241T variant and the RAD51-G135C homozygote-type significantly increase the risk of the development of AML with inv(16)/t(16;16)/CBFβ-MYH11.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Child ; Child, Preschool ; DNA-Binding Proteins ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Leukemia, Myeloid, Acute ; etiology ; genetics ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Polymorphism, Single Nucleotide ; Rad51 Recombinase ; genetics ; Translocation, Genetic ; Young Adult

OBJECTIVETo investigate the implications of erythroblasts periodic acid-Schiff (PAS) stain for myelodysplastic syndromes (MDS) dyserythropoiesis, diagnosis and differential diagnosis.

METHODSPAS stain of bone marrow (BM) erythroblasts in 406 MDS patients, 207 non-severe aplastic anemia (NSAA), 144 immune thrombocytopenic purpura (ITP), 67 megaloblastic anemia (MegA), 76 iron deficiency anemia (IDA), 50 paroxysmal nocturnal hemoglobinuria (PNH), and 50 acute erythroid leukemia (AEL) as well as some related laboratory parameters in MDS patients were analyzed retrospectively.

RESULTSPAS-positive detection rate was significantly higher in MDS (53.0%) than in NSAA (14.5%), ITP (27.1%) and PNH (16.0%), but was significantly lower in MDS than in AEL (84.0%) (all P = 0.000). There was no significant difference in PAS-positive detection between MDS and MegA (46.3%), or MDS and IDA (40.8%) (P = 0.310, 0.052, respectively). Erythroblasts PAS-positive rate (Median, M = 1%) and PAS-positive scores (M' = 2) was significantly lower in MDS than in AEL (M = 8%; M' = 17), and significantly higher than in NSAA (M = 0%; M' = 0), ITP (M = 0%; M' = 0), PNH (M = 0%; M' = 0), MegA (M = 0%; M' = 0), and IDA (M = 0%; M' = 0) (all P < 0.05). The cut-off value of PAS-positive rate and score for distinguishing MDS from the other groups except AEL were 0.5% and 0.5, with a sensitivity and specificity of 60.8% and 74.4%, respectively. For MDS patients, the percentage of BM erythroid cells was significantly higher in PAS-positive group than in PAS-negative group (P < 0.05), and so were megakaryocyte count, lymphocyte-like micromegakaryocytes count and percentage of micromegakaryocyte (P = 0.002, 0.000, 0.000, respectively). HGB, MCV, MCH and MCHC were significantly lower in PAS-positive group (all P < 0.05), and so was the neutrophil alkaling phosphatase (NALP) (P = 0.000). PAS-positive detection rate, positive rate and score were higher in MDS patients with abnormal karyotype than with normal karyotype, and were also higher in IPSS high/intermediate-risk 2 group than in low/intermidiate-risk 1 group.

CONCLUSIONThe positive reaction of erythroblasts PAS stain is an indicator of dyserythropoiesis. It is helpful to the diagnosis of MDS patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Diagnosis, Differential ; Erythroblasts ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; Periodic Acid-Schiff Reaction ; Retrospective Studies ; Sensitivity and Specificity ; Staining and Labeling ; Young Adult

Up to now, no consensus has been reached on the standard salvage regimen for patients with refractory or relapsed acute myeloid leukemia (AML). This study was purposed to evaluate the efficacy and safety of combination chemotherapy composing of cyclophosphamide (Cy), cytosine arabinoside (Ara-C) and topotecan (CAT regimen) for 37 refractory or relapsed AML patients. The dosing regimen was as follows: Cy 300 mg/m2 by intravenous infusion, every 12 hours on days 1-3, topotecan 1.25 mg/m2 by intravenous continuous infusion over 6 hours daily on days 2 to 6, Ara-C 500 mg/m2 by intravenous infusion over 2 hours daily for 5 days on days 2-6. The results showed that all patients completed one cycle of chemotherapy. 12 patients (32.4%) achieved complete remission (CR), 2 (5.4%) achieved partial remission (PR), and the 23 remaining patients achieved no remission (NR). The overall response rate (RR) was 37.8%. Among 18 relapsed cases, 6 cases had CR (33.3%), 2 cases achieved PR (11.1%), and 10 cases were with NR (55.6%). Among 19 refractory cases, 6 had CR (31.6%), and 13 (68.4%) were with NR. There was no statistically significant difference in RR between refractory and relapsed groups (31.6% and 44.4%, respectively) (p=0.42). Myelosuppression was universal. Mild non-hematologic toxicities were mainly gastrointestinal, as nausea, vomiting, diarrhea. The incidence of severe (grade III-IV) non-hematologic toxicity, such as oral mucositis and infection was 37.8% and 86.5% respectively. Only one patient died of severe infection during the observation (within 28 days from start of chemotherapy). The time of median follow-up was 4 (0-33) months, the median overall survival (OS) was 4 (1.8-6.2) months. The median OS for responders was longer than that for non-responders (9 vs 2 months respectively, p=0.00). In conclusion, the CAT regimen of lower dose is well tolerated and has certain anti-leukemia effect, and worthy to be further investigated.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; administration & dosage ; Cytarabine ; administration & dosage ; Female ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Male ; Middle Aged ; Recurrence ; Topotecan ; administration & dosage ; Young Adult