OBJECTIVETo explore the relationship between microRNA and drug resistance in leukemia treatment by screening and identifying the microRNAs which differentially express in K562 cell line and its adriamycin resistant cells-K562/A02 cell line.

METHODSThe drug resistance potency of K562/A02 cells was evaluated by MTT assay. P-gp expression of K562 and K562/A02 cells were detected by flow cytometry (FCM). The differentially expressed microRNAs in K562 and K562/A02 cells were analyzed by microarray technique and Real Time RT-PCR.

RESULTSThe resistance to adriamycin (ADM) of K562/A02 cells was 180 fold greater than that of K562 cells. P-gp expression rate of K562 and K562/A02 cells was 0.2% and 86%, respectively. Twenty-two microRNAs expressed differentially in K562 and K562/A02 cells (P < 0.01). As compared to K562 cells, expressions of miR-221, miR-155 and miR-451 were up-regulated by more than two fold, while expression of miR-98, miR-181a, let-7f, let-7g, miR-424 and miR-563 down-regulated by more than two fold in K562/A02 cells. The results of real time RT-PCR were consistent with that of microarray. Of note, differential expressions of miR-451, miR-155, miR-221, let-7f and miR-424 were remarkable.

CONCLUSIONK562/A02 cells show a different microRNA expression profile as compared to its parental K562 cells, suggesting microRNAs including miR-221, miR-155, miR-451, let-7f and miR-424 may be involved in the mechanism of drug resistance in leukemia. These differentially expressed microRNAs provide potential novel targets for overcoming drug-resistance.

Doxorubicin ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; MicroRNAs ; genetics

OBJECTIVETo study the plasma HCII activity and antigen level variations and their relationship with arterial and deep venous thrombotic diseases.

METHODSSeventy-five patients with brain infarction (BI), 50 myocardial infarction (MI), 36 deep venous thromboembolic disease (DVT) and 50 healthy controls were entered in this study. Plasma HCII activity was measured with chromogenic substrate method and the HCII antigen level by Western blotting assay. Plasma antithrombin (AT) activity was detected for the HCII deficiency individuals with DVT using chromogenic substrate method.

RESULTSThere was no significant difference in the mean plasma HCII activity and antigen levels between BI group [(99.97 +/- 21.14)% and 0.96 +/- 0.24], MI group [(98.18 +/- 29.35)% and 0.95 +/- 0.20] and healthy controls [(96.80 +/- 20.11)% and 0.93 +/- 0.19]. The plasma HCII activity and antigen concentrations in patients with DVT [(89.57 +/- 17.12)% and 0.87 +/- 10.18] tended to be decreased as compared with healthy controls, but they were not significant. No significant difference was found for the prevalence of HCII deficiency between patient groups and control group. The HCII deficiency individuals with DVT had normal AT activity and fibrinogen concentration.

CONCLUSIONSPlasma HCII deficiency may not be the risk factor for arterial thrombosis in the Han population of Hunan Chinese. It is needed to further confirm if decreased plasma HCII is correlated with venous thrombosis.

Adult ; Aged ; Blotting, Western ; Cerebral Infarction ; blood ; etiology ; Female ; Heparin Cofactor II ; analysis ; deficiency ; immunology ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; etiology ; Venous Thrombosis ; blood ; etiology

OBJECTIVETo screen phosphopeptide specific for acute leukemia.

METHODSMononuclear cells from bone marrow were collected from 16 newly diagnosed acute lymphoblastic leukemia (ALL) and 20 acute myeloid leukemia (AML) patients. Peptides were extracted and purified, analyzed by immunoprecipitation and liquid chromatography coupled with tandem mass spectrometry (LC-MS).

RESULTS(1) Non-receptor tyrosine kinase family members Fyn, Yes, Src widely expressed in acute leukemia; (2) Some phosphopeptides, including non-receptor tyrosine kinase family members Abl/iso1 and Abl, non-receptor Ser/Thr protein kinase family members Bcr, JNK2, JNK2 iso2, Adaptor/scaffold members Cas-L, Cbl, CrkL CENTD1 (Centaurin delta1) ZO2, transcriptor GFR-1 and phosphatase SHIP-2 were detected in Ph positive ALL, but not in other kinds of ALL. (3) Hck, Lyn and Fgr selectively expressed in AML (except AML-M(3)).

CONCLUSIONSome phosphopeptides were specific for ALL and AML, and may be useful for diagnosis and therapy of acute leukemia.

Chromatography, High Pressure Liquid ; Humans ; Immunoprecipitation ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm Proteins ; analysis ; Phosphopeptides ; analysis ; Phosphorylation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Proteomics ; Tandem Mass Spectrometry

BACKGROUNDA nonsteroidal anti-inflammatory drug, indomethacin, has been shown to have anti-leukemic activity and induce leukemic cell apoptosis. This study was to elucidate the mechanism of indomethacin-induced K562 cell apoptosis.

METHODSK562 cells were grown in RPMI 1640 medium and treated with different doses of indomethacin (0 micromol/L, 100 micromol/L, 200 micromol/L, 400 micromol/L, 800 micromol/L) for 72 hours. The cells were harvested, and cell viability or apoptosis was analyzed using MTT assay and AO/EB stain, combining laser scanning confocal microscopy (LSCM) technique separately. For the localization and distribution of intracellular caspase-3 or caspase-8 protein, immunofluorescence assay was carried out. To reveal the activation of caspase-3 or caspase-8 in indomethacin-treated cells, Western blot detection was used. The change in intracellular free calcium was determined by Fluo-3/Am probe labeling combined with LSCM.

RESULTSIndomethacin could lead to K562 cell apoptosis and inhibit cell viability in a concentration-dependent manner. An increased expression of intracellular caspase-3 or caspase-8 was observed at higher doses of indomethacin (400 - 800 micromol/L). Western blot results showed upregulation and activation in both caspase-3 and caspase-8 protein. Under indomethacin intervention, the levels of intracellular free calcium showed a significant increase. Blocking the activity of cyclooxygenase did not abolish the effects of indomethacin on K562 cell apoptosis.

CONCLUSIONSActivation and upregulation of caspase-3 or caspase-8 protein were responsible for Indomethacin-induced K562 cell apoptosis. Variation of intracellular free calcium might switch on the apoptotic pathway and the proapoptotic effect of indomethacin might be cyclooxygenase-independent.

Apoptosis ; drug effects ; Calcium ; metabolism ; Caspase 3 ; Caspase 8 ; Caspases ; genetics ; metabolism ; Cyclooxygenase Inhibitors ; pharmacology ; Enzyme Activation ; Gene Expression Regulation, Enzymologic ; drug effects ; Humans ; Indomethacin ; pharmacology ; K562 Cells

OBJECTIVETo monitor the changes of hemolysis parameters and endothelial cell markers in thrombotic thrombocytopenic purpura (TTP) and reveal the clinical significance of these changes.

METHODSvWF-cleaving protease (vWF-CP) activity in 3 cases of TTP was detected by Western blot. The percentages of fragmented red cells (FRC) were counted throughout the entire clinical course. Levels of plasma thrombomodulin were detected by Western blot combined with density screening in TTP and healthy individuals (n = 3). Concentration of plasma VEGF was measured by enzyme-linked immunosorbent assay in TTP and healthy individuals (n = 9). Fundus fluorescein angiography was performed to search the evidence of microvascular thrombosis in one TTP patient with impaired visual acuity.

RESULTSThe lower vWF-CP activity was observed in TTP patients; the percentages of FRC in 3 cases of TTP were 1.65%, 2.50%, 3.32% respectively with an average of 2.49% at the onset of and decreased with the improvement of the disease. The levels of plasma TM and VEGF were significantly elevated in TTP than those in healthy individuals, and related to the severity of TTP. Fundus photography in one TTP patient with impaired visual acuity revealed vascular occlusion in fundus arteriole and venulae.

CONCLUSIONSA decreased vWF-CP activity is in favour of TTP diagnosis. Dynamic monitoring of plasma TM and VEGF as well as percentages of FRC are useful indexes for reflecting the severity and evaluating therapeutic response of TTP. Selective fundus fluorescein angiography is useful for the judgement of microvascular thrombosis in TTP.

ADAM Proteins ; blood ; ADAMTS13 Protein ; Biomarkers ; blood ; Blotting, Western ; Endothelial Cells ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Hemolysis ; Humans ; Male ; Middle Aged ; Purpura, Thrombotic Thrombocytopenic ; blood ; pathology ; Thrombomodulin ; blood ; Time Factors ; Vascular Endothelial Growth Factor A ; blood ; Young Adult

OBJECTIVE: To understand the clinical features and histopathology of histocytic necrotizing lymphadenitis (HNL) so as to better recognize the disease. METHODS: The clinical features, histopathology, and diagnosis of 10 patients admitted to our hospital were retrospectively analyzed. RESULTS: The clinical features of these 10 cases included: young females were the majority; lymphadenopathy and fever were the most common clinical manifestations; some cases were accompanied by connective tissue diseases. Histopathologic examination showed distinctive necrosis and around the necrotic foci, variable proliferations of histocytes but generally without infiltration of neutrophils. CONCLUSION HNL has some typical histopathological alterations and relatively fine prognosis,but it tends to be misdiagnosed as lymphoma or lymphoid tuberculosis and may be accompanied by other diseases.
Adolescent ; Adult ; Age Factors ; Diagnosis, Differential ; Female ; Histiocytic Necrotizing Lymphadenitis ; diagnosis ; pathology ; Humans ; Lymph Nodes ; pathology ; Lymphoma ; diagnosis ; Male ; Middle Aged ; Retrospective Studies ; Sex Factors ; Tuberculosis, Lymph Node ; diagnosis

OBJECTIVETo investigate the effect and mechanism of BSDW on the model of allergic rhinitis and the model of guinea pigs by histamine shocking in guinea pigs.

METHODUsing the model of allergic rhinitis in guinea pigs caused by 10% TDI, we observed the effect of BSDW on physiological and pathological symptoms of allergic rhinitis in guinea pigs, the effect of the levels of serum IgE and serum and nasal histamine. Using the model of guinea pigs by histamine shocking, we observed the effect of BSDW on physiological symptoms in guinea pigs.

RESULTBSDW significantly relieved the pathological symptoms of allergic rhinitis in guinea pigs, alleviated the hyperplasia of columnar epithelium, decreased the number of monocyte and eosinocyte compared with the model group. It also reduced the levels of serum IgE, and decreased the release of serum and nasal histamine. BSDW significantly prolonged the occurent time of gasping, eclampsia and death caused by shock, reduced the times of gasping in the model of guinea pigs by histamine shocking.

CONCLUSIONBSDW has significant effect against allergy. The mechanism relates to its effects of decreasing the levels of serum IgE and inhibiting the release of serum and nasal histamine.

Administration, Intranasal ; Animals ; Anti-Allergic Agents ; pharmacology ; Asarum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Guinea Pigs ; Histamine ; blood ; Immunoglobulin E ; blood ; Lamiaceae ; chemistry ; Male ; Nasal Mucosa ; immunology ; Plants, Medicinal ; chemistry ; Rhinitis, Allergic, Perennial ; immunology ; Scutellaria ; chemistry ; Toluene 2,4-Diisocyanate

To prepare Fe(3)O(4)-magnetic nanoparticles loaded with adriamycin and investigate the reversal role of drug-loaded nanoparticles in K562 and resistant cell line K562/A02, the drug-loaded nanoparticles were prepared by using mechanical absorption polymerization at different conditions of 4 degrees C or 37 degrees C for 24 or 48 hours. The survival of cells cultured with drug-loaded nanoparticles for 48 hours was detected by MTT assay, then the growth inhibition efficacy of cells was calculated. The results showed that the growth inhibition efficacy of both two cell lines was enhanced with increasing concentration of Fe(3)O(4)-magnetic nanoparticles. The inhibitory ratio of two cell lines obtained at 4 degrees C and for 48 hours was significantly better than that at 37 degrees C and 24 hours. In conclusion, Fe(3)O(4)-magnetic nanoparticles can load adriamycin by using mechanical absorption polymerization, but depended on proper temperature and time. Furthermore, drug-loaded nanoparticles showed an ability reversing multidrug resistance.
Doxorubicin ; chemistry ; pharmacology ; Drug Carriers ; chemistry ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Ferric Compounds ; pharmacology ; Humans ; K562 Cells ; Magnetics ; Nanoparticles ; chemistry ; Particle Size

Adult ; Aged ; Angioplasty, Balloon, Coronary ; Diabetes Complications ; etiology ; Female ; Humans ; Male ; Middle Aged ; Stents ; Treatment Outcome

OBJECTIVETo compare the procedural and in-hospital outcomes in a large series of diabetic and non-diabetic patients undergoing selective percutaneous coronary intervention (PCI) and to evaluate the influence of diabetes mellitus on the procedural and in-hospital outcomes.

METHODS1294 consecutive patients underwent selective PCI from January to December 2002 in this institution were analyzed retrospectively. Baseline clinical, in-lab and in-hospital outcome information were recorded. Rates of procedural success, device success and clinical success were analyzed and logistic regression was performed to model the association between diabetes status and outcomes.

RESULTSTwo hundred and sixty-nine patients (20.8%) complicated with diabetes. Type C lesion, double and triple vessel diseases were more prevalent in diabetics than those in non-diabetics. The pre-PCI diameter stenosis of diabetics was significantly more severe than that of non-diabetics (91.00 +/- 6.62 vs 89.81 +/- 6.64, P < 0.01). The balloon length, maximum balloon diameter and maximum balloon inflation pressure, maximum inflation duration were larger in diabetics than those in non-diabetics [(17.07 +/- 6.31) mm vs (16.07 +/- 7.28) mm, (2.30 +/- 1.11) mm vs (2.12 +/- 0.94) mm, (9.86 +/- 4.40) atm vs (9.05 +/- 4.75) atm, (20.94 +/- 14.69) s vs (18.26 +/- 14.65) s, respectively, P < 0.05]. The stent diameter was smaller in diabetics than that in non-diabetics [(3.15 +/- 0.47) mm vs (3.23 +/- 0.43) mm, P < 0.05]. The procedural success rate showed no significant difference between two groups (89.6% vs 90.3%, P > 0.05). But a higher incidence of acute/subacute stent thrombosis was observed in diabetics compared with that in non-diabetics (1.9% vs 0.5%, P < 0.05). The rate of clinical success was similar between diabetics and non-diabetics (99.3% vs 99.2%, P > 0.05). Diabetes was not an independent predictor of acute outcomes in the regression model.

CONCLUSIONSA higher incidence of acute/subacute stent thrombosis was observed in diabetics. The incidence of procedural and in-hospital major adverse cardiac events and the rate of clinical success were similar between diabetics and non-diabetics. Diabetes was not an independent predictor of in-hospital outcomes after selective PCI.

Aged ; Angioplasty, Balloon, Coronary ; Coronary Stenosis ; complications ; therapy ; Diabetes Mellitus, Type 2 ; complications ; Drug-Eluting Stents ; Female ; Humans ; Length of Stay ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome