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Objective To assess the clinical significance of detecting the immune markers in idiopathic thrombocytopenic purpura (ITP). Methods The frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombopoietin in 64 ITP patients and 31 healthy controls were measured with enzyme-linked immunospot assay (ELISPOT),modified monoclonal antibody immunobilization of platelet antigens assay (MAIPA), flow cytometry and sandwich enzyme-linked innnunosorbent assay respectively. Results Compared with the controls[1.3 ± 0. 5/105 peripheral blood mononuclear cell (PBMC), (0.33±0.06,0.41±0.03), (22.08±4.54)% and (8.19±2.46)%], the frequencies of circulating B cells secreting platelet-specific antibody (7.6±4.6/105 PBMC in acute ITP group, 5.3±3.0/105 PBMC in chronic ITP group), platelet-specific antibody (including the anti-GP Ⅱ b/Ⅲa antibody, anti-GP Ⅰ b/X antibody) (0.51 ±0.11, 0.48±0.06 in acute ITP group; 0.49±0.10,0.46±0.09 in chronic ITP group), the percentage of CD8+ T Lymphocyte (27.09±9.86 ) %, the percentage of reticulated platelet in ITP patients[the megakaryocyte cytosis group (24. 85 ± 19. 18)%, the normal megakaryocyte group (23.89±18.90)%]were significantly increased ( all P＜0.05).The frequencies of circulating B cells secreting platelet-specific antibody in acute ITP patients were notably increased (P＜0.05) compared to the chronic ITP patients. In T lymphocyte subsets, the percentage of CD3+T lymphocyte and CD4+ T lymphocyte and the ratio of CD4+/CD8+ in the patients with ITP[(60.88±14.59)%, (28.41±10.55)%, 1.18±0.59]were notably decreased than those in the healthy controls [(69.89±6.43)%, (35.38±5.05) %, 1.64±0.29, P＜0.05]. There was no apparent difference of the level of thrombopoietin between ITP patients with megakaryocyte cytosis (72. 09 ± 41.64 ) and health controls (75.37± 26. 32, P ＞ 0. 05 ), however, the level of thrombopoietin of ITP patients with normal megakaryocyte apparently increased (118.60±70.72, P＜0.05). Conclusion Detecting the frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombepoietin in the patients with ITP may improve the diagnosis and guide clinical therapy.
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Objective To analyze clonal chromosomal abnormalities of patients with MDS and evaluate prognosis combining with complete blood counts and blast counts. Methods The chromosomes were prepared with direct method, brief culture of cells and R-banding techniques, and then the karyotypie analysis was performed. Results Abnormal chromosomes were detected 44.9%, including the numeral abnormalities of chromosomes and structural alterations.The most common chromosomal aberrations were -7,+8.The rate of abnormal karotype in refractory anemia with erythroblasts (RAEB1 and RAEB2) was much higher than in refractory anemia (RA) and refractory anemia with ringed sideroblasts (RAILS). Chromosomal abnormalities correlated positively with bone marrow blasts. Conclusion Karyotype analysis is very useful for the diagnosis, treatment and prognosis evaluation in MDS.
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Objective To comprehensively evaluate the curative effect and adverse effects of rituximab plus cyclophosphamide, vincristine, doxoruhicin and prednisone(R-CHOP) chemotherapy and CHOP chemotherapy alone on the treatment for low and moderate malignant B cell non-Hodgkin lymphoma (NHL). Methods By the application of the systematic review method of Cochrance International Collaboration, the world-wide randomized controlled trials (RCT) on the comparison between different curative effects of R-CHOP and CHOP chemotherapy alone on the treatment low and moderate malignant B cell NHL was collected and the study results was evaluated systematically. Results Seven RCT studies involving 1569 patients and had no heterogeneity between themselves (χ=5.31,P=0.50). The baseline of patients characteristics was comparable in all the studies. By comparing complete response (CR) rate and adverse effects through fixed effect analysis model, the results showed that R-CHOP was better than CHOP chemotherapy on the treatment for low and moderate malignant B cell NHL(OR =2.22, 95 %CI 1.72-2.85, P ＜0.000 01), and adverse events of R-CHOP had no significant difference compared with CHOP chemotherapy alone (P＞0.05). Conclusion With good curative effect on the treatment low and moderate malignant B cell NHL and without obvious differences from the CHOP chemotherapy alone in adverse effects, R-CHOP chemotherapy should be recommended as the best treatment method for low and moderate malignant B cell NHL And much more well-designed clinical RCT should be made to further prove its clinical effect.
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Objective To study the influences of the length of sexual abstinence on sperm parameters and seminal plasma biochemical indicator.Methods Sperm concentration and motility were detected by computer assisted sperm analysis(CASA).The sperm morphology was analyzed using semiautomatic sperm morphology analyzer.Seminal plasma fructose concentration,neutral a-glycosidase,zinc,and acid phosphates concentration were measured by spectrophotometer.Improved DTNB was conducted for detection of the content of seminal plasma novain.Prostatespecific antigen(PSA)was measured with commercial kit.Three groups were defined according to the days of sexual abstinence:G1(1-3 days),G2(4-5 days)and G3(＞6 days).Results Sperm concentration of G2 was significantly higher than that of G1(P＜0.01),and of G3 than that of G1 and G2(P＜0.01).Sperm motility and per centage of morphologically normal sperm of G3 were significantly lower than that of G1(P＜0.01).Seminal plasma fructose concentration of G3 was significantly lower than that of G1 and G2(P＜0.01,P＜0.05).Seminal plasma neutral a-glycosidase,carnutine,zinc concentration of G3 was significantly higher than that of G1 and G2(P＜0.01,P＜0.05).Conclusion The length of sexual abstinence can influence sperm parameters and seminal plasma biochemical indicator.
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By preventing mesenchymal stem cells (MSCs) from adhering to precoated agarose to create a model of MSC suspension in vitro, we investigated anoikis in MSCs and the role of caspase-3 in the anoikis. The cultured MSCs were randomly divided into 3 groups: the anoikis group,caspase-3 inhibitor group and control group. Before experiment, we coated dishes with 1.5 % agarose;in the anoikis group, MSCs were put into the precoated dishes; and in the inhibitor group, caspase-3 inhibitor and MSCs were also put into the precoated dishes; but there were not intervention in the control group. MSCs were collected at 2 h, 6 h, 12 h and 24 h. The alteration of caspase-3 activity was evaluated by caspase-3 fluorometric assay and western blot analysis. The apoptosis rates were detected by flow cytometry. MSCs were round and suspended sufficiently in the anoikis and inhibitor groups. Caspase-3 fluorometric assay showed that there were significant differences in statistics between the anoikis group and the others (P＜0.05). Western blot analysis discovered that caspase-3 expression.in the anoikis group was more than that in the control and inhibitor groups (P＜0.05). Flow cytometry showed that the apoptosis peak appeared in all the three groups, but it increased dramatically in the anoikis group. The apoptosis rates in the inhibitor and control groups were low and stable.And there were significant differences in statistics between the anoikis group and the others (P＜0.05).MSCs will undergo anoikis in suspended condition if they are separated from the extracellular matrix.Caspase-3 inhibitors can suppress caspase-3 activity and reduce the apoptosis rate significantly. Caspase-3 plays a vital part in induced MSC anoikis in vitro. MSCs suspension culture system might be set up with argorose and caspase-3 inhibitor.
ABSTRACT
By preventing mesenchymal stem cells (MSCs) from adhering to precoated agarose to create a model of MSC suspension in vitro, we investigated anoikis in MSCs and the role of caspase-3 in the anoikis. The cultured MSCs were randomly divided into 3 groups: the anoikis group, caspase-3 inhibitor group and control group. Before experiment, we coated dishes with 1.5 % agarose; in the anoikis group, MSCs were put into the precoated dishes; and in the inhibitor group, caspase-3 inhibitor and MSCs were also put into the precoated dishes; but there were not intervention in the control group. MSCs were collected at 2 h, 6 h, 12 h and 24 h. The alteration of caspase-3 activity was evaluated by caspase-3 fluorometric assay and western blot analysis. The apoptosis rates were detected by flow cytometry. MSCs were round and suspended sufficiently in the anoikis and inhibitor groups. Caspase-3 fluorometric assay showed that there were significant differences in statistics between the anoikis group and the others (P<0.05). Western blot analysis discovered that caspase-3 expression in the anoikis group was more than that in the control and inhibitor groups (P<0.05). Flow cytometry showed that the apoptosis peak appeared in all the three groups, but it increased dramatically in the anoikis group. The apoptosis rates in the inhibitor and control groups were low and stable. And there were significant differences in statistics between the anoikis group and the others (P<0.05). MSCs will undergo anoikis in suspended condition if they are separated from the extracellular matrix. Caspase-3 inhibitors can suppress caspase-3 activity and reduce the apoptosis rate significantly. Caspase-3 plays a vital part in induced MSC anoikis in vitro. MSCs suspension culture system might be set up with argorose and caspase-3 inhibitor.
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The present study consisted of the assay of the POA activity (SGPT. SGOT and SMAO) in 20 cases of cirrhosis of liver. Similar study was also done in CCl4 rabbit model Pathological examination was carried out for the animal livers.By comparing the clinical observations of patients with the results of experimental animal we could see that the estimation of POA activity might reflect the degree of liver cell degeneration, necrosis and fibrosis of cirrhotic patients, and they could also be used as a biochemical index for evaluation of the therapeutic effect. It was considered further that the prognosis of cirrhosis could be predicted if serum aldolase isoenzyme A (and/ or serum AFP level) activity was estimated in addition.