OBJECTIVETo measure the subsets of dendritic cells 1 (DC1) in the bone marrow of severe aplastic anemia (SAA) patients and evaluate the relationships between the CD11c+CD83+ cells and Th1 cells, CD3+CD8+ cells or hematopoietic function and explore the role of DC1 in the pathogenesis of SAA.

METHODSBy FACS, the quantities and ratios of CD11c+CD1a+ cells, CD11c+CD83+ cells, Th1 cells, and CD3+CD8+ cells in the bone marrow of SAA patients and normal controls were detected respectively. The relationships between CD3+CD8+ cells and reticulocyte absolute value (Ret) or neutrophil absolute value (ANC), between Th1 cells and CD3+CD8+ cells, Ret or ANC, between CD11c+CD83+ cells, and Th1 cells, CD3+CD8+ cells, Ret or ANC were evaluated.

RESULTSIn normal controls' bone marrow, the percentages of Th1 cells, CD11c+CD1a+ cells, CD11c+CD83+ cells and the ratio of CD11c+CD83+/CD11c+CD1a+ were (0.42 +/- 0.30)%, (0.38 +/- 0.29)%, (0.37 +/- 0.32)% and 1.07 +/- 0.10, respectively. In untreated SAA patients, they were (4.87 +/- 0.54)%, (1.73 +/- 0.24)%, (3.38 +/- 0.56)% and 2.21 +/- 0.32 respectively, which were higher than that in normal controls (P < 0.01). In recovering SAA patients, the percentages of Th1 cells, CD11c+CD1a+ cells and CD11c+CD83+ cells decreased significantly to (0.53 +/- 0.22)%, (0.61 +/- 0.23)%, (0.65 +/- 0.22)%, respectively (P < 0.01). The ratio of CD11c+CD83+/CD11c+ CD1a+ in recovering SAA patients decreased to 1.37 +/- 0.25, which was similar to that in normal controls (P > 0.05). The percentage of CD3+CD8+ cells in untreated SAA patients was (32.32 +/- 10.22)%, and in recovering SAA patients decreased to (13.67 +/- 5.24)% (P < 0.01). The percentage of CD3+CD8+ cells in SAA patients was negatively correlated with their Ret and ANC (P < 0.05), while their Th1 cell percentages were positively correlated with their CD3+CD8+ cells (P < 0.01), and negatively correlated with their Ret and ANC (P < 0.01). SAA patient's CD11c+CD83+ cell percentages were positively correlated with their Th1 cell and CD3+CD8 cells (P < 0.01, P < 0.05), but negatively with their Ret and ANC (P < 0.01).

CONCLUSIONBoth immature DC1 and activated DC1 increased in the bone marrow of SAA patients, and the balance of DC1 subsets shifted from stable form to active one, which might promote Th0 cells to polarize to Th1 cells, and cause the over-function of T lymphocytes and hematopoiesis failure in SAA.

Adolescent ; Adult ; Anemia, Aplastic ; immunology ; Antigens, CD ; immunology ; Antigens, CD1 ; immunology ; Bone Marrow ; immunology ; CD11c Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Dendritic Cells ; immunology ; Female ; Humans ; Immunoglobulins ; immunology ; Male ; Membrane Glycoproteins ; immunology ; Th1 Cells ; immunology ; Young Adult

This study was aimed to investigate the effect of homoharringtonine (HHT) on K562 cell proliferation, apoptosis and expression of BCL-2 and NF-κB proteins. The cells proliferation was assayed with MTT method, the cell apoptosis, cell cycle and BCL-2 expression were analyzed with flow cytometry, NF-κB protein expression was detected with Western blot. The results showed that HHT concentration-dependently inhibited proliferation of K562 cells, the IC50 at 48 h was 43.89 ng/ml. Treated with HHT 10 ng/ml for 48 h, K562 cell apoptosis significantly increased, cell cycle was blocked at G0/G1, the expression level of BCL-2 and NF-κB proteins was lower than that in control group (P < 0.05). It is concluded that HHT may inhibit the proliferation of K562 cells, and down-regulating expression levels of BCL-2 and NF-κB may be one of its anti-CML mechanisms.
Flow Cytometry ; Harringtonines ; pharmacology ; Humans ; K562 Cells ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

Type Ⅶ secretion system(T7SS) is a novel and specialized secretion system discovered in recent years. It was first found in Mycobacterium tuberculosis. Type Ⅶ secretion system is involved in the secretion of virulence-associated proteins, the interaction between pathogens and hosts and the balance of zinc/iron ions. Moreover,it plays a critical role in the growth and pathogenesis of Mycobacteria. This review summarizes the components,substrates and translocation mechanisms of the type Ⅶ secretion system and its relation to the virulence of Mycobacteria aiming to provide references for developing novel strategies for disea-ses diagnosis,treatment and prevention.

Objective To study the mechanism of two-component system of MprAB and TrcRS in synergistically regulating gene rv1057 expression of Mycobacterium tuberculosis.Methods The in vivo specific binding capability of MprA and TrcR with the target gene promoter region was analyzed using electrophoretic mobility shift assay.The transcription level of target gene was analyzed by using fluorescence quantitative polymerase chain reaction,and all results were compared with the fold changes in H37Rv strain plus SDS group,which was set as one unit.The expression level of target gene was analyzed by using western blot;the transcription ability of different promoter region of rv1057 was detected through lacZ report gene.The t test was used for statistical analysis.Results MprA was able to bind to trcR promoter.The expressions of trcR in D981 and H37Rv strains without SDS were 1.7 and 2.5 folds of the expression of H37Rv strains with SDS groups,respectively.The difference between these two groups was statistically significant (t=18.54,P＜0.05).With SDS,the expressions of trcR in D981 and H37Rv strains were 1.0 and 2.1 folds of the expression of H37Rv strains plus SDS group,respectively.The expressions of trcR in D981 and H37Rv strains were significantly different (t=15.86,P＜0.05).After adding SDS during the culture of H37Rv strains,the expression of trcR in H37Rv decreased.The difference between these two groups was statistically significant (t=16.99,P＜0.05).Both MprA and TrcR were able to bind to rv1057 promoter and regulate its expression.MprA activated the expression of rv1057,while TrcR repressed the expression of rv1057.Conclusions MprAB and TrcRS synergistically regulate the expression of rv1057.MprA is activated in the presence of SDS,which represses the transcription of trcR and activates the transcription of rv1057.However,TrcR represses the transcription of rv1057 in the absence of SDS.

OBJECTIVETo isolate Japanese encephalitis virus (JEV) from mosquitoes collected in Tanghe County, Henan province and analyze the genotype of the newly isolated JEV strains and the characteristics of amino acid in the E gene.

METHODSViruses were isolated from mosquitoes collected in 2004 and identified by biological, serological and molecular biologic methods. PrM and E segments of the newly isolated JEV were amplified by RT-PCR, the PCR products were purified and sequenced. Multiple alignment, phylogenetic and amino acid (AA) analysis were carried out by Clustal X (1.8) program, MEGA 3.1 and GENEDOS (3.2).

RESULTSTotally 3722 mosquitoes were collected including Culex, Armigeres, Aedes, Anopheline. Three new JEV strains isolated from Culex belonged to genotype 1. The homologue of nucleotide and amino acid of E gene between new JEV strains and live attenuated vaccine strain SA14-14-2 was 86.9-87.7% and 95.2%-97.0%, respectively. Totally there were 12 common sites of amino acid differences in E gene between them.

CONCLUSIONNewly isolated viruses in Henan province belonged to JEV genotype 1. It suggests that the vaccine strain SA14-14-2 currently used for preventing JE is able to protect people from JEV infection, although there are some amino acid differences between them.

Animals ; Animals, Newborn ; Cell Line ; China ; Culicidae ; virology ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; blood ; virology ; Genotype ; Insect Vectors ; virology ; Mice ; Phylogeny ; RNA, Viral ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Amino Acid ; Sequence Homology, Nucleic Acid

OBJECTIVETo compare the treatment results of three-dimensional conformal radiotherapy (3D-CRT) and conventional radiotherapy (2D) for patients with locally advanced non-small-cell lung cancer (NSCLC).

METHODSFive hundred and twenty seven patients with stage III NSCLC treated between Jan 2000 and Dec 2006 were included in this study. Among them, 253 cases were treated with 3D-CRT, and 274 with conventional radiotherapy. In the 3D group, 159 (62.8%) patients received chemoradiotherapy, 77 with total radiotherapy dose of > 60 Gy, 49 with 50 - 60 Gy. In the 2D group, 127 (46.4%) patients received chemoradiotherapy, 48 with total radiotherapy dose of > 60 Gy, 75 with 50 - 60 Gy.

RESULTSThe 1-, 3-, 5-year overall survival rates (OS) and median survival time for patients treated with 3D-CRT were 73.3%, 26.1%, 14.4% and 20.1 months, respectively, and that of patients treated with 2D radiotherapy were 61.0%, 13.8%, 8.0% and 15.6 months, respectively (P = 0.002). The 1-, 3-, 5-year cause-specific survival rates (CSS) were 79.0%, 33.3%, and 20.8% for the 3D group and 65.1%, 16.7%, 11.2%, respectively, for the 2D group (P = 0.000). The 1-, 3-, and 5-year locoregional control rates were 71.6%, 34.3% and 31.0% for patients treated with 3D radiotherapy and 57.3%, 22.1% and 19.2%, respectively, for patients treated with 2D treatment (P = 0.002). The results of multivariate analysis showed that 3D-CRT, KPS, clinical tumor response and pretreatment hemoglobin level were independently associated with increased OS and CSS. No statistically significant differences were found between the radiation complications in the two groups.

CONCLUSIONSThe results of our study demonstrate that 3D-conformal radiotherapy improves the survival rate in patients with stage III NSCLC compared with that of 2D radiation therapy.

Aged ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; radiotherapy ; Chemoradiotherapy ; Female ; Follow-Up Studies ; Hemoglobins ; metabolism ; Humans ; Lung Neoplasms ; drug therapy ; pathology ; radiotherapy ; Male ; Neoplasm Staging ; Radiation Pneumonitis ; etiology ; Radiotherapy Dosage ; Radiotherapy, Conformal ; adverse effects ; methods ; Survival Rate

OBJECTIVETo observe the efficacy and side effect of DA/HA regimen chemotherapy for the treatment of refractory and relapsed paroxysmal nocturnal hemoglobinuria (PNH).

METHODSEight patients with refractory and relapsed PNH were treated with DA/HA regimen chemotherapy. Three patients were treated with DA (DNR 40 mg/d, i.v.drip, the first and the second day; 20 mg/d, i.v.drip, the third day; Ara-C 100 mg/d, i.v.drip, for 5 days) and 5 patients with HA (HHT 2 - 3 mg/d, i.v.drip, for 5 days; Ara-C 100 mg/d, i.v.drip, for 5 days).

RESULTSAll the 8 patients responded well: the PNH clone was diminished in five patients. Hemolysis was remitted in 6 cases. Five patients showed improvement in hematological parameters. The dosage of corticosteroid was decreased in all of them. No serious side effect was revealed.

CONCLUSIONDA/HA regimen chemotherapy was safe and effective for refractory and relapsed PNH patients.

Adolescent ; Adult ; Cytarabine ; administration & dosage ; Daunorubicin ; administration & dosage ; Drug Therapy, Combination ; Female ; Glycosylphosphatidylinositols ; analysis ; Harringtonines ; Hemoglobinuria, Paroxysmal ; drug therapy ; Humans ; Male

OBJECTIVETo analyse the relapse rate and risk factors of autoimmune hemolytic anemia (AIHA) and Evans syndrome.

METHODSFifty two cases of AIHA and Evans syndrome in remission being followed up for 1 - 14 years (median time 3.8 years) were analysed for relapse rate. The risk factors of relapse were analysed by case-control study.

RESULTSThe total relapse rate of these AIHA and Evans syndrome patients was 57.7%, and the median remission duration to the first relapse was 9 months. The relapse rates in patients with negative Coombs test, warm autoantibodies and both of warm and cold autoantibodies were 30.8% (4/13), 54.0% (13/24) and 86.7% (13/15), respectively. The relapse rate in patients with cold antibody was the highest (P < 0.05). The relapse rate in patients with antibody titer >or= 100 was 92.9% (13/14) and was higher than that in patients with antibody titer < 100 [59.5% (13/22)] (P < 0.05). Patients treated with prednisone and cyclosporin relapsed less than those treated with prednisone alone, and the relapse was related to the therapy course of prednisone and CsA.

CONCLUSIONBecause of the high relapse rate, AIHA and Evans syndrome should be treated according to the class of autoantibodies, and with longer course of prednisone and cyclosporin and prophylaxis of infection.

Adolescent ; Adult ; Aged ; Anemia, Hemolytic, Autoimmune ; etiology ; immunology ; Autoantibodies ; blood ; Child ; Cyclosporine ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Recurrence ; Risk Factors ; Syndrome ; Thrombocytopenia ; etiology ; immunology

OBJECTIVETo study the clinical features of severe aplastic anemia (SAA) patients with complication of infection.

METHODSA retrospective analysis of prevalence of infection occurring in 229 SAA patients, their bacterial spectrum, and the effect of GM-CSF or G-CSF on the infection were done.

RESULTThe prevalence of infection in SAA patients was 86.0%, among which 54.2% was infected with gram-positive organisms, 40.0% with gram-negative bacilli and 5.8% with fungal infections. Septicemia occurred mostly with E. coli and Pseudomonas infection. Patient's neutropenia was significantly related to the infection. The patients with neutrophil count less than 0.2 x 10(9)/L had more frequent and severe infection. Age, hemoglobin level, subtype of T lymphocytes and antithymocyte globulin therapy were not related to infection. Prophylaxis usage of floxacin could not reduce patient' gastrointestinal infection. The total mortality of SAA patients with infection was 23.1%. Pulmonary infection and septicemia increased mortality, and GM-CSF/G-CSF therapy reduce mortality.

CONCLUSIONSAA patients were at high risk of infection which was significantly associated with severe neutropenia. GM-CSF or G-CSF therapy exerts an assistant role to antibiotics in controlling the infections.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; complications ; Anti-Bacterial Agents ; therapeutic use ; Bacteria ; isolation & purification ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infection ; drug therapy ; etiology ; Male ; Middle Aged ; Time Factors

This study was aimed to evaluate expression levels of CD166, Fas and apoptosis-related proteins in bone marrow neutrophils of PNH patients and normal controls, and to analyze their correlation in order to explore whether exist apoptosis abnormality in BM neutrophils of PNH patients. The expression levels of CD16b, Fas and Bax, Bcl-2 in BM neutrophils of PNH patients and normal controls were assayed by flow cytometry; the difference of expression levels between patients and controls, and expression correlation between CD16b and apoptosis-related proteins were compared. The results showed that (1) the expression levels of CD16b on BM neutrophils of patients and controls were (20.36 +/- 9.05)% and (71.34 +/- 26.8)% respectively (P = 0.01); (2) the expression levels of CD95 on BM neutrophils of patients and controls were (62.83 +/- 32.11)% and (48.00 +/- 38.52)% respectively, there were no significant difference between CD95 expressions in BM neutrophils of PNH patients and controls and no significant correlation between expression of CD95 and CD16b on BM neutrophils of PNH patients (P > 0.05); (3) the expression levels of Bcl-2 in BM neutrophil cytoplasma of patients and controls were (8.64 +/- 5.40)% and (16.82 +/- 15.39)% respectively, there were no significant difference between Bcl-2 expression of patients and controls, and no significant correlation between the expression of Bcl-2 and CD16b in BM neutrophil cytoplasma of PNH patients (P > 0.05); (4) the expression levels of Bax in BM neutrophil cytoplasma of patients and control were (30.47 +/- 22.15)% and (48.47 +/- 15.99)% respectively, there were no significant difference between the Bax expressions of patients and controls, and no significant correlation between the Bax and CD16b expressions in BM neutrophil cytoplasma of PNH patients. In conclusion, BM neutrophils of PNH patients expressed apoptosis-related CD95, Bcl-2 and Bax without significant difference from the normal controls, and without significant correlation with the CD16b expression. It is suggested that the cell growth and decrease of PNH patients possibly are independent of abnormal apoptosis.
Adolescent ; Adult ; Apoptosis Regulatory Proteins ; biosynthesis ; Bone Marrow Cells ; metabolism ; pathology ; Female ; Flow Cytometry ; GPI-Linked Proteins ; Hemoglobinuria, Paroxysmal ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Neutrophils ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Receptors, IgG ; biosynthesis ; bcl-2-Associated X Protein ; biosynthesis ; fas Receptor ; biosynthesis