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.

Purpose:To investigate the effect of MMF on the postoperative infection after renal transplanta-tion. Methods: Before 1997, the CsA + Aza + prednisone were used as immunosuppresive agents postoperativelyfor 56 cases; Since 1998, drugs used for 34 cases were changed into MMF and CsA+MMF+prednisone respec-tively. The dose initially used for CsA was 65 mg/kg · d 1 in the patients of Aza group and 5 mg/kg · d 1 for theMMF group and then this dosage was regulated according to the determination of plasma level of the drug. Re-sults:The total infection rate of these 90 patients was 17.8%, 12 cases were infected in the Aza group (21.4%)and 11.8% (4 cases) in the MMF group. There were 2 patients died in the Aza group. Conclusions:The postop-erative infection rate of the patients underwent kidney transplantation might be reduced when MMF was used tosubstitute Aza during the period of postoperative management.

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 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

C57BL/6 mice were inoculated intranasally (50 microl) with serial 10-fold dilution of HAB/01 H5N1 virus. Three and five days later, three mice of each group were euthanized. Lung injury was assessed by observation of lung histopathology, virus titers and MCD50 were also measured. Our data showed that H5N1 viral infection in mice resulted in mainly epithelial injury and interstitial pneumonia, featuring significant weight loss, dramatically increased lung wet weight:body weight ratio, inflammatory cellular infiltration, alveolar and interstitial edema, hemorrhage in lungs with high virus titers, and MCD50 was 10(-6.5)/ 0.05 mL. These results suggested that a mouse model of H5N1 viral infection was successfully established which may benefit study of H5N1 avian influenza virus and pathogenic mechanism of host.
Animals ; Brain ; pathology ; virology ; Disease Models, Animal ; Female ; Humans ; Influenza A Virus, H5N1 Subtype ; pathogenicity ; Influenza, Human ; pathology ; virology ; Liver ; pathology ; virology ; Lung ; pathology ; virology ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Spleen ; pathology ; virology

OBJECTIVETo analyze the clinical features and prognosis of the primary myelodysplastic syndrome with myelofibrosis (MDS-MF) patients and to improve the cognition of MDS-MF.

METHODSFour hundred and sixty-six primary MDS patients with bone marrow (BM) biopsy were divided into two groups according to whether BM associated with fibrosis, the clinical features and prognosis of the two groups were analyzed retrospectively.

RESULTS167 (35.8%) MDS cases revealed myelofibrosis, of which MF-1 123 cases (26.4%), MF-2 40 cases (8.6%), MF-3 4 cases (0.9%). The proportion of hepatosplenomegaly in MDS-MF group was significantly higher than in MDS without MF group, the difference had statistical significance (P = 0.031). The proliferation of BM biopsy in MDS-MF group was significantly more active than in MDS without MF group. The number of blasts, megakaryocytes and abnormal megakaryocytes in MDS-MF group were significantly higher than in MDS without MF group, the differences had statistical significance (P < 0.05). Among the 345 patients who had available results of cytogenetic analysis, 121 cases were MDS-MF patients, the proportion of middle and high-risk prognostic group according to IPSS karyotype prognosis groups in MDS-MF group were significantly higher than in MDS without MF group, the differences had statistical significance (P = 0.047). The median survival was 17 (1 - 60) months in MDS-MF group, and was 32 (1 - 62) months in MDS without MF group. The difference had statistical significance (P = 0.001). Myelofibrosis had independent prognostic significance by multi-variable analysis (P = 0.019).

CONCLUSIONThe myelofibrosis in MDS is main the proliferation of reticular fiber. The proliferation of reticular fiber is closely related with the number of blast cells, the proliferation and developmental abnormalities of megakaryocytes and the karyotype. The prognosis of MDS-MF patients is poor.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; diagnosis ; pathology ; Primary Myelofibrosis ; complications ; diagnosis ; pathology ; Prognosis ; Retrospective Studies ; Young Adult