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Background: Studies have shown that aromatherapy improve health problems related to anxiety, depression, heart rate variability (HRV), and sleep. However, the effect of aromatherapy in women who suffer from menopausal syndrome and its specific effects on HRV and sleep quality are unknown., Purpose: This study designed an aromatherapy intervention and evaluated its effect on menopausal syndrome, HRV, and sleep quality in women., Methods: This double-blinded randomized controlled trial was conducted at a medical center hospital. A total of 84 participants who met the study criteria were randomly assigned using permuted block randomization. The experimental group received a 20-min inhalation of essential oil and the control group received a 20-min inhalation of sweet almond oil every night for 4 weeks. Posttest data was collected at 2 weeks after completion of the intervention. Data were collected using the Green Menopausal Symptom Scale, HRV device, and Pittsburgh Sleep Quality Index., Results: After adjusting for age, the results of the generalized estimation equations (GEE) showed that all outcomes were significantly different in both the experimental group and the control group (p < .05) for the interaction effect of group and time, and that the outcome of sleep quality on the 2nd week in the control group was not significantly different (p < .066)., Conclusions / Implications for Practice: This study supports that the 4-week aromatherapy intervention improves menopausal symptoms, the activity of autonomic nervous systems, and sleep quality in women with no adverse side effects. In the future, this intervention may be applied in outpatient departments to promote the health of menopausal women.

Objective: To investigate the effects of synthetic long-chain polyphosphate on blood coagulation and platelet aggregation., Methods: The effect of artificial synthetic long chain poly phosphate on blood coagulation and platelet aggregation was detected by coagulation tests, coagulation factor activity detection and platelet aggregation test, and its mechanism was explored by ELISA, flow cytometry and high content imaging system., Results: The long chain polyphosphates prolonged activated partial thromboplastin time, decreased coagulation factor FⅧ, FⅨ, FⅪ and FⅫ activity, blocked ADP-induced platelet aggregation, and decreased the concentration of calcium and TXA2 in platelet., Conclusion: The synthetic long-chain polyphosphate can inhibit endogenous coagulation and inhibit platelet aggregation, which may be related with the inhibition of intracellular calcium and TXA2.

Since the American Medical Association published the 2011 guidelines for immune thrombocytopenia, China has been the first to update the guidelines for immune thrombocytopenia based on evidence-based medicine. Recently, there have been many breakthroughs in clinical research published, especially the Chinese medical workers have made a prominent contribution to the treatment of the immune thrombocytopenia. However, the references of systematic drug introduction for children, adults, aged and pregnant women are still insufficient, and the first or second line treatment for some patients were ineffective. Therefore, we tried to combine the references to interpret the guidelines, to explore the advantages and disadvantages of each treatment, to find out the bottleneck of clinical treatment, so as to facilitate the implementation and understanding of the guidelines, then update the next guideline.

Objective: To investigate the efficacy and safety of IA regimen which contains idarubicin (IDA) 8 mg/m(2), 10 mg/m(2) or 12 mg/m(2) as induction chemotherapy for adult patients with de-novo acute myeloid leukemia (AML) . Methods: A total of 1 215 newly diagnosed adult AML patients, ranging from May 2011 to March 2015 in the First Affiliated Hospital of Soochow University and other 36 clinical blood centers in China were enrolled in the multicenter, single-blind, non-randomized, clinical controlled study. To compare the response rate of complete remission (CR) , adverse events between different dose idarubicin combined with cytarabine (100 mg/m(2)) as induction chemotherapy in newly diagnosed patients of adult AML. Results: Of 1 207 evaluable AML patients were assigned to this analysis of CR rate. The CR rates of IDA 8 mg/m(2) group, IDA 10 mg/m(2) group and IDA 12 mg/m(2) group were 73.6% (215/292) , 84.1% (662/787) and 86.7% (111/128) , respectively ( P <0.001) . After adjusted for age, blast ratio of bone marrow, FAB classification and risk stratification, the odds ratios (95% CI ) of IDA 10 mg/m(2) group and IDA 12 mg/m(2) group were 0.49 (0.34-0.70) and 0.36 (0.18-0.71) , as compared with the IDA 8 mg/m(2) group ( P <0.001, P =0.003) . In the intermediate and favorable groups, CR rates was 76.5% (163/213) , 86.9% (506/582) and 86.1% (68/79) in different doses of IDA ( P =0.007) . Interestingly, IA regimen with IDA 10 mg/m(2) was the only beneficial factor affecting CR in this group after adjusted for age, blast ratio of bone marrow and FAB classification[ OR =0.47 (95% CI 0.31-0.71) , P <0.001]. CR rates in adverse group was 50.0% (18/36) , 60.6% (43/71) and 81.8% (18/22) respectively ( P =0.089) . However, the odds ratios (95% CI ) of IDA 12 mg/m(2) when compared with the IDA 8 mg/m(2) was 0.22 (0.06-0.80) , after adjusted for age, blast ratio of bone marrow and FAB classification. The median time (days) of neutrophil count less than 0.5×10(9)/L in IDA 8 mg/m(2) group, IDA 10 mg/m(2) group and IDA 12 mg/m(2) group were 14 (11-18) , 15 (11-20) and 18 (14-22) , respectively ( P =0.012) and of platelet count lower than 20×10(9)/L were 14 (7-17) , 15 (11-20) and 17 (15-21) , respectively ( P =0.001) . The incidences of lung infection in the three groups were 9.8%, 13.5% and 25.2%, respectively ( P <0.001) . Conclusions: For young adult patients (aged 18-60 years) with AML in China, intensifying induction therapy with idarubicin 10 mg/m(2) is clinically superior to IDA 8 mg/m(2) and IDA 12 mg/m(2) in favorable intermediate AML subgroup. However, idarubicin 12 mg/m(2) is more suitable to adverse AML subgroup.

Objective: To evaluate the efficacy and safety of maintenance therapy with reduced dose of rhTPO in the patients with primary immune thrombocytopenia (ITP) who attained stable platelet (PLT) counts after daily administration of rhTPO. Methods: Treatment was started with a daily administration of rhTPO (300 U/kg) for 2 consecutive weeks. Patients who attained stable PLT≥50×10(9)/L were enrolled to maintenance therapy starting with every other day administration of rhTPO, then adjusted dose interval to maintain platelet count (30-100) ×10(9)/L. Results: A total of 91 eligible patients were enrolled. Fourteen patients discontinued the study due to noncompliance (12/14) and investigator decision (2/14) . Among 77 patients who completed the study, 38 patients with the administration of rhTPO at every other day or less could maintain PLT≥30×10(9)/L for 12 weeks. The percentage of patients with a platelet response (PLT≥30×10(9)/L) at 4(th) week, 8(th) week and 12(th) week of maintain therapy was 92.6% (63/68) , 82.7% (43/52) and 85.0% (34/40) , respectively. Median platelet counts remained in the range of (70-124) ×10(9)/L. The overall incidence of rhTPO-related adverse events was 7.7%. All the adverse events were generally mild. Conclusion: Extending the dose interval of rhTPO is feasible to maintain stable platelet count in the patients with ITP, but the optimal dose interval is uncertain and might vary with individuals.

Objective: To find the risk loci on cholecystokinin A receptor gene (CCKAR) - a schizophrenia candidate gene by using the deep sequencing and then analyze the variations., Methods: In the present study, 8 schizophrenia patients and 8 healthy controls were recruited. After DNA extraction from peripheral blood, we conducted deep sequencing on CCKAR region by HaloPlex Target Enrichment System (Agilent). We used Unphased software to exclude the false positive., Results: After deep sequencing, we got 103 loci, among which 30 were located in CCKAR gene. Besides, the SNP rs191275118 was found to be associated with schizophrenia., Conclusions: A new variation that may be associated with schizophrenia was found. The deep sequencing is effective to find genetic variation.

Objective: To screen and analyze CD34(+) cell specific microRNAs (miRNAs) from the patients with acute myelogenous leukemia (AML) and their expression., Methods: CD34(+) cells were sorted from AML patients or the mobilized peripheral blood of the donors of hematopoietic stem cell transplantation (normal control subjects) and followed by the extraction of the cell total RNAs. The differentially expressed microRNAs (miRNAs, miR) were selected after hybridizing with miRNA microarray, real time polymerase chain reaction (real-time PCR) was subsequently applied to confirm the expression of the selected miRs, and PCR products were further cloned and sequenced to check their specificity., Results: Of the differentially expressed miRNAs, 191 were found to be at least one-fold change in the CD34(+) cells between the AML patients and the normal control subjects. Of the 191 miRNAs, the expression difference of 94 was significant (P < 0.05). Among these 94 miRNAs, the expression of 44 miRNAs was increased and the other 50 miRNAs was decreased in the CD34(+) cells from the bone marrow of AML patients compared with the CD34(+) cells from the mobilized peripheral blood of the normal control subjects. Real time PCR verified that the expression level of miR-10a and miR-220c in the CD34(+) cells from the bone marrow of AML patients was 19.6% and 19.0% of that of CD34(+) cells from mobilized peripheral blood of the normal control subjects. DNA sequencing and BLAST DNA database searching results indicated that the PCR products were really miR-10a and miR-220c., Conclusion: A variety of differentially expressed-miRNAs are existed between AML and normal control subjects CD34(+) cells, the expression of miR-10a and miR-220c was significantly down-regulated in the CD34(+) cells from the bone marrow of AML patients.

A little is known about the specific marker on the surface of acute leukemia cells, leading to the lack of the specific diagnosis method for acute leukemia. Therefore, in this study, cell-systematic evolution of ligands by exponential enrichment (cSELEX) was performed to screen the aptamers binding to CD33(+)/CD34(+) cells from the patients with acute myeloblastic leukemia (AML) of M(2) subtype (AML-M₂) so as to provide the basis for finding the specific marker on the surface of AML-M(2) CD33(+)/CD34(+) cells. Firstly, AML-M₂ CD33(+)/CD34(+) cells were sorted and used as targeted cells, and normal CD33(+)/CD34(+)cells were used as counter-targeted cells; the aptamers binding to CD33(+)/CD34(+) cells from patients with AML-M₂ were screened from the single strand deoxyribonucleic acid (ssDNA) library by cSELEX. Subsequently, each aptamer structure was analyzed after cloning and sequencing. The results indicated that after 13 round of screenings, the enrichment of aptamers in the ssDNA library was ranged from 0.7% to 52.9%, and reached steady state at 13th round screening. Sequence analysis for 30 aptamers showed that most of the aptamers born one of the three conserved sequences of CCCCT, CTCTC, and CTCAC. Secondary structure analysis indicated that three different secondary structures existed in these aptamers. It is concluded that the aptamers binding to the AML-M(2) CD33(+)/CD34(+) cells are successfully screened, which lay the basis for further looking for the specific marker on the surface of AML-M₂ CD33(+)/CD34(+) cells, and the molecular diagnosis of the AML-M₂ leukemia.

Constitutive activation of nuclear transcription factor-κB (NF-κB) exists in a variety of leukemia, and induction of apoptosis through blocking NF-κB activation may be an alternative strategy for leukemia treatment. The aim of this study was to investigate the inducing effect of modified adenovirus 5-based adenovirus vector (i.e. chimeric Ad5F35 Vec)-mediated expression of mutant IκBα (IκBαDN) on apoptosis of HL-60 cells. The recombinant Ad5F35-IκBαDN Vec carrying IκBαDN cDNA which deleted the first 1-70 amino acids coding sequences at 5' terminal of human IκBα was transfected into HL-60 cells. The apoptosis, NF-κB DNA binding activity, the expressions of IκBα, cIAP-2 and xIAP in HL-60 cells were detected by DNA binding assay, flow cytometry, real-time quantitative polymerase chain reaction and Western blot respectively. The results showed that apoptosis rates were 22.53 ± 2.999%, 6.08 ± 2.464% and 4.86 ± 1.366% for Ad5F35-IκBαDN Vec-infected or blank vector of Ad5F35-EGFP Vec-transfected and untransfected HL-60 cells respectively, which showed a significant difference between Ad5F35-IκBαDN Vec-transfected and untransfected cells (p < 0.001) and between Ad5F35-IκBαDN Vec-transfected and Ad5F35-EGFP Vec-transfected cells (p < 0.001, p < 0.002), while NF-κB DNA binding activity was decreased, the truncated IκBα was expressed, and IκBα mRNA expression was up-regulated, but the expression of cIAP-2 and xIAP mRNA was down-regulated after transduction for 48 hours. It is concluded that the chimeric Ad5F35 Vec can effectively mediate the expression of IκBαDN cDNA in HL-60 cells, leading to the inhibition of NF-κB DNA binding activity and inducing apoptosis of HL-60 cells.

The aim of this study was to investigate the effect of 3-O-acetyl-11-keto-β-boswellic acid (AKBA) on the proliferation, apoptosis and cell cycle of human acute myeloid leukemia (AML) cell line HL-60. HL-60 cells were treated by AKBA at various concentrations. The inhibitory effects of AKBA on the proliferation of HL-60 were analyzed by MTT assay. Morphologic changes of HL-60 cells were observed by fluorescence microscopy with Hochest33342 staining. Cell apoptosis rate was determined by flow cytometry with Annexin-V-FITC/PI double staining. The cell cycle was measured by flow cytometry with PI staining. The results showed that AKBA inhibited the proliferation of HL-60 and the apoptosis rate of HL-60 cells was gradually enhanced when AKBA dose increased. AKBA changed the cell cycle of HL-60, resulting in cell increase at G(1) phase and decrease at S phase. It is concluded that the AKBA has anti-proliferation and apoptosis-inducing effects on HL-60 cells, that seems a promising therapeutical approach for AML.

This study was aimed to explore the expression level of angiotensin-II type 1 receptor (AT1) mRNA in bone marrow of myeloid leukemic patients, and its correlation with the proportion of leukemia cells in samples and Hb, WBC, Plt counting in peripheral blood. 51 samples, including 36 AML, 7 CML, and 8 samples of non-malignant hematological diseases as control group were collected. The expression of at1 mRNA was detected by real time-PCR; the expression levels of at1 gene in AML and CML groups were relatively quantitatively analyzed by using 2(-ΔΔCT) and were compared with control group. The results showed that the expression levels of at1 mRNA in AML, CML and control groups were 0.038 ± 0.076, 0.033 ± 0.039, 0.281 ± 0.366, respectively. at1 gene expression in the myeloid leukemic group was significantly lower than that in the control group. The expression level of at1 mRNA in AML was negatively correlated with the proportion of leukemia cells and positively with Hb level in peripheral blood. It is concluded that at1 gene may play a minor role in leukaemogenesis, however, may promote erythropoiesis.

Objective: To investigate the effect of Dahuangzhechong pill on the gene expression spectra of preventing arterial thrombosis, and reveal its mechanism on molecule level., Methods: Mononuclear cell and blood platelet of the arterial thrombosis patients were separated before and after treatment by Dahuangzhechong pill. Their RNA was extracted respectively and the genes expressions were detected using gene array containing 14,000 gene., Results: 44 genes up-expressed and 299 genes down-expressed in blood platelet, 252 genes expression increased and 299 genes expression decreased in mononuclear cell genes after treated with Dahuangzhechong pill. The cluster analysis showed that the genes contained ion channel and transport protein, apoptosis related protein, DNA synthesis, repair and transcription factor, cell receptor, cell signal and transducin, and protein translation and synthesis, etc., Conclusion: Dahuangzhechong pill may prevent arterial thrombosis through genes containing ion channel and transport protein, apoptosis related protein, DNA synthesis, repair and transcription factor, cell receptor, cell signal and transducin, and protein translation and synthesis, etc.

Objective: To detect the levels of serum amyloid A (SAA) and explore the pathogenesis of cardiovascular diseases in patients with obstructive sleep apnea syndrome (OSAS)., Methods: Polysomnography was performed in 80 patients with OSAS and 20 control subjects matched for age and body mass index. The patients with OSAS were divided into mild OSAS group (n=22), moderate OSAS group (n=23) and severe OSAS group (n=35) according to the apnea hypopnea index (AHI). Serum amyloid A levels were measured in all the subjects by enzyme-linked immunosorbent assay for correlation analysis., Results: Serum amyloid A levels in mild OSAS group (1.66-/+0.73 microg/ml), moderate OSAS group (2.72-/+1.12 microg/ml) and severe OSAS group (4.08-/+1.85 microg/ml) were significantly higher than those in the control group (0.66-/+0.59 microg/ml) (P<0.05). SAA levels also differed significantly between the 3 OSAS groups (P<0.01), increasing with the severity of OSAS. Correlation analysis indicated that SAA level was positively correlated to AHI (r=0.649, P<0.01) and TSaO(2)<90% (r=0.491, P<0.01), but inversely yo miniSaO(2) (r=-0.499, P<0.01). After 3 months of nCPAP therapy, SAA levels were significantly decreased in the 20 patients with severe OSAS (4.13-/+2.27 microg/ml vs 5.14-/+2.30 microg/ml, P<0.01)., Conclusion: SAA levels are elevated in OSAS patients in close correlation to the severity of OSAS, which may contribute to the vulnerability of the patients to cardiovascular diseases. nCPAP therapy help reduce the risk for cardiovascular diseases in OSAS patients.

Objective: To establish and evaluate a rabbit model of arterial thrombosis by modified thread-drawing., Methods: Fifty-three rabbits were randomly divided into 6 groups: a normal group, a ligating group,a collagen encapsulated thread-drawing group,a aspirin group,a clopidogrel group, and an aspirin clopidogral group. The endovascular pathological changes in the rabbits were observed, and D-fructose-1,6-diphosphate trisodium salt octahydrate (FDP), D-Dimer and tissue factor (TF) were detected with enzyme linked immunosorbent assay., Results: In the thread-drawing group, thrombus was obvious, and the endovascular elastic membrane was injured seriously compared with the ligating group. After being treated with aspirin and clopidogrel, most arterial thrombus was softened, dissolved and absorbed. Compared with that in the modified thread-drawing group,wet and dry weight of thrombus increased,and the level of D-Dimer, FDP and TF also increased in the modified thread-drawing group (P<0.01). After being treated by aspirin and/or clopidogrel, the wet and dry weight of thrombus and the level of D-Dimer, FDP and TF decreased compared with the control (P<0.01). Aspirin plus clopidogral could obviously reduce the wet and dry weight of thrombus, and reduce the level of D-Dimer and FDP (P<0.01). Aspirin plus clopidogral could obviously inhibit the formation of TF compared with aspirin (P<0.05)., Conclusion: Arterial thrombosis model by collagen encapsulated thread-drawing which is visible, repeatable and effective is better than thread-drawing. It is suitable for screening anti-thrombosis drugs and evaluating their effect.

Objective: To evaluate the clinical efficacy and toxicity of priming induction regimen of CAG for newly diagnosed acute myeloid leukemia (AML) in elderly patients., Methods: Seventy-five patients with newly diagnosed AML were divided into 2 groups: 34 were treated with priming induction regimen CAG and the other 41 were treated with 2 classic routine chemotherapy regimens including pirarubicin+cytarabine (TA) and homoharringtonine+cytarabine (HA). All patients had a 14 day interval between the 2 courses of chemotherapy., Results: The complete remission rate after 2 courses of induction therapy in patients with the priming induction regimen CAG and the total efficacy rate was significantly higher than that of the routine chemotherapy patients(67.6% vs. 39%; 82.4% vs. 56.1%). Patients with unfavorable karyotypes had poor chemotherapy efficacy. The 3-year disease-free-survival (DFS) time was longer in patients with AML treated with priming induction regimen CAG than in patients treated with 2 classic routine chemotherapy regimens. Except for the muscular soreness, the hematological and non-hematological side effects in the CAG priming induction group were significantly fewer than those in the routine chemotherapy group., Conclusion: The priming induction regimen of CAG has a significantly higher complete remission rate and an efficacy rate, fewer side effects, milder chemotherapy intensity and is more sensitive to chemotherapeutic drugs than those of the routine chemotherapy. It can shorten the duration of agranulocytosis and decrease infectious complications and increase the sensitivity of leukemia blast cells to chemotherapeutic drugs.

Objective: To explore the molecular mechanism of Glanzmann thrombasthenia (GT)., Methods: All 45 exons of alphaIIb and beta3 subunit genes as well as their splicing sites were amplified by polymerase chain reaction(PCR) with 40 primer pairs, and then the PCR products were used to screen the gene mutation by single strand conformation polymorphism-polyacrylamide gel electrophoresis (SSCP-PAGE). The mutation was further confirmed by direct DNA sequencing., Results: A DNA band alterated migration was detected after SSCP-PAGE. DNA sequencing showed that a base deletion within the band at the site of 540 in GPIIb gene(540A) was found., Conclusion: The frame-shift mutation caused by the deletion of 540A in GPIIb gene is a novel mutation which is a genetic defect in patients with GT.

Objective: To establish a duplex TaqMan RT-PCR assay for urine DD3/PSA mRNA ratio, and to evaluate its effect in diagnosis of prostate cancer (PCa)., Methods: Urine samples were obtained from 34 patients with PCa and 44 patients with benign prostate hypertrophy (BPH) by prostate massage. A duplex TaqMan RT-PCR assay was developed: PCR primers were designed to amplify the fragments between the exon1 and exon2 in the PSA mRNA and between the exon1 and exon3 in the DD3 mRNA, and the PCR TaqMan-MGB probes were labeled with HEX and FAM respectively in 5' for PSA mRNA. LNCaP cells were used as template. DD3/PSA mRNA ratio was measured. Receiver operating characteristic curve (ROC) was drawn so as to evaluate its diagnostic efficacy., Results: Sequencing showed that the PCR products were specific for PSA mRNA and DD3 mRNA. The minimum detection level was approximately 0.6 cells/reaction for PSA mRNA and was 60 cells/reaction for DD3 mRNA in the LNCaP cell cDNA. The intra- and inter-assay coefficients of variation of DD3/PSA mRNA were 3.8%-4.7% and 4.1%-4.9% respectively. Urine DD3/PSA mRNA ratio in PCa group was significantly higher than the BPH group (P < 0.01). When the cutoff value was defined as 0.254, the area under curve (AUC) of DD3/PSA mRNA ratio was 0.746 (95% CI: 0.630-0.862), and the sensitivity and specificity were 64.7% and 77.3% respectively. The urine DD3/PSA mRNA ratio positive rate was not correlated with clinical and pathological parameters., Conclusion: A duplex TaqMan RT-PCR assay for urine DD3/PSA mRNA ratio has been established with an excellent clinical performance and specificity for PCa, saving time and reducing costs. It may be a promising method in the early diagnosis of PCa.

Objective: To examine the expression of the inhibitor alpha of nuclear transcription factor kappaB (IkappaBalpha) mRNA expression and its sequence characteristics in human nasopharyngeal carcinoma cell (NPC) lines CNE1, CNE2, HNE1 and HNE2., Methods: Reverse transcription was performed with the total RNAs isolated from the NPC cell lines CNE1, CNE2, HNE1 and HNE2, as well as the transplanted tumor tissues with HNE1 cells. Then IkappaBalpha cDNA was amplified by PCR, and the products were used to examine IkappaB alpha mRNA expression and DNA sequencing, or the DNA sequencing after the products were cloned into plasmid vector., Results: IkappaB alpha mRNA was expressed in all the 4 nasopharyngeal carcinoma cell lines. DNA sequencing showed that polymorphisms and 5 mutations (A825G, A975G, G576A, A655G and C653A) existed in IkappaB cDNA from the transplanted tumor tissues with HNE1 cells, CNE1 and CNE2 cells., Conclusion: The expression of IkappaBalpha mRNA not only exists, but DNA polymorphisms and some additional mutations in IkappaBalpha cDNA are also detected in the nasopharyngeal carcinoma cells.

Objective: To study the effects of insulin like growth factor-1 (IGF-1) on cell viability and tissue factor (TF) in angiotensin II (Ang II) induced vascular endothelial cells and to investigate its mechanisms., Methods: 10(-6) mol/L Ang II was added to human vascular endothelial cells (HUVECs) culture media alone or 30 min after pretreatment with IGF-1 (0.1 microg/ml , 0.5 microg/ml, 2.5 microg/ml). Cell viability and AngII type 1 receptor (AT1-R) mRNA were evaluated after 24 h incubation with AngII. At the optimum concentration of IGF-1 affecting cell viability, the time dependent manner for 12 - 48 h incubation with Ang II was evaluated. TF, NOS and NO were investigated after 24 h incubation with Ang II. In addition, NO synthase inhibitor Nomega-nitro-1-arginine methylester(L-NAME) was added 30 min before addition of IGF-1 and Ang II, and cell viability, TF, AT1-R mRNA, NOS and NO were evaluated after 24 h incubation., Results: (1) Ang II induced a decrease in cell vitality, an upregulation of AT1-R mRNA, an increase in TF, and a decrease in the activity of NOS and content of NO. (2) Pretreatment with IGF-1 significantly inhibited the decreased cell viability and upregulation of AT1-R mRNA. IGF-1 at 0.5 microg/ml showed the most obvious effects. This effect of cell viability recovery was in a time dependent manner during 12 -48 h. (3) IGF-1 also inhibited the increased content of TF, the decreased activity of NOS and the decreased content of NO. (4) The beneficial effects of IGF-1 on cultured endothelial cells were completely abolished by L-NAME., Conclusion: IGF-1 pretreatment could enhance the ang II injured cell viability and anti-thrombosis capacity, and the protective effects may be related to activation of NOS-NO signaling pathway which inhibited AT1-R.

Objective: To investigate the relationship among 3 polymorphisms of GP IIb and the function of GP IIb T13959 G in the platelet transfusion refractoriness(PTR)., Methods: The 26th exon, the 30th exon and the 21st intron of gene GP IIb in 110 individuals were amplified by polymerase chain reaction (PCR), and the PCR products were analyzed with single-strand conformation polymorphism(SSCP) and sequenced to investigate whether there was linkage among the polymorphisms of the gene. Human platelet antigen-3 (HPA-3) gene frequency was detected by Fok I enzyme in 147 patients with hematologic diseases, and was compared with that in 110 normal individuals. Forty-four patients who received apheresis platelet transfusion repeatedly were randomly divided into the HPA-3 homotype group and the control group. The antibodies of the platelet were detected after 3 times of platelet transfusion., Results: There were polymorphisms of gene GP IIb in the 26th, 30th exon and the 21st intron, and the mutations were: T changed into G in 13,959 th of the 26th exon; C changed into T in 16,997 th of the 30th exon; the 9 bps deletion occurred in 11,996-12,004 th of the 21st intron. The 3 polymorphisms had synchronization in the individuals. The results of Fok I enzyme indicated that the frequency of HPA-3a was 83.6% (92/110)and 81.9%(119/147), and that of HPA-3b was 16.4%(18/110) and 19.1%(28/147) in the normal individuals and the patients respectively. There was no significant difference between the patients and normal individuals (P>0.05). After the platelet transfusion, the antibodies of all the cases of the homotype platelet transfusion were negative, but the antibodies in 2 cases of the control group were positive, and there was antibody to HPA-3a in one of the antibodies positive cases., Conclusion: (1)There is close linkage among the polymorphisms of gene GP IIb, which is T->G in 13 959 th of the 26th exon, C->T in 16,997 th of the 30th exon, and the 9 bps deletion in 11,996-12,004 th in the 21st intron. (2)The gene frequency of HPA-3a/3b is similar in the normal individuals and patients with hematologic diseases. (3) HPA-3 system may be one of the reasons for PTR in Chinese.

Objective: To determine the effects of Tongxinluo on cell viability and tissue factor (TF) in AngII induced vascular endothelial cells and to investigate its mechanism., Methods: AngII(10(-6)mol/L) was added to human vascular endothelial cells (HUVECs) culture media alone or with various concentration of Tongxinluo drug containing plasma (5%,10%, and 20%) added 30 minutes before AngII. Cell viability was evaluated after 24-hour incubation with AngII in a dose manner. TF, AngII type 1 receptor (AT(1)) mRNA, NO synthase (NOS) and NO were observed after 24-hour incubation with AngII. In addition, NOS inhibitor nomega-nitro-larginine (L-NAME) was added 30 minutes before Tongxinluo and AngII. Cell viability, TF, AT(1)mRNA, the level of NOS and NO were evaluated after 24-hour incubation with Tongxinluo and AngII., Results: Tongxinluo significantly improved AngII induced endothelial cell viability and the effect was the most obvious at 10%. Tongxinluo (10%) decreased the TF and AT(1) mRNA while increased the NOS and NO levels. L-NAME obviously inhibited the effects of Tongxinluo on cell viability, TF, AT(1) mRNA, and NOS and NO levels., Conclusion: Up-regulating NOS-NO signaling may be the mechanism of Tongxinluo on cell viability and TF in AngII induced vacular endothelial cells.

Objective: To explore the effect and toxicity profile of recombinant human interleukin 11(rhIL-11) on the platelet after hematopoietic stem cell transplantation in patients with leukemia., Methods: Twenty-four patients with acute or chronic leukemia treated by allogeneic peripheral blood stem cell transplantation (PBSCT) were randomly divided into a test group and a control group. The patients in the test group were treated with rhIL-11 since the 13th day after PBSCT (1.5mg/d),while the control group were given symptomatic treatment., Results: The average time for the platelet to recover to the level of 20 x 10(9)/L was 20.8 days in the test group, and 26.0 days in control group respectively, there was significant difference (P<0.01). The average time for the platelet to recover to the level of 50 x 10(9)/L was 25.7 days in the test group, and 32.3 days in the control group respectively, there was also significant difference (P<0.01). The average time for the platelet transfusion was 2.2 in the test group, 4.1 in the control group, and there was significantly different (P<0.01). The average number of megakaryocytes was 12.2 in the test group, 4.8 in the control group on 30th day after the transplantation,and there was significant difference(P<0.01). The main side effects of rhIL-11 were nausea, vomit, debility, headache, dizzy and pain of injection site, and the degree was all Iapproximately II grade., Conclusion: rhIL-11 has definite recuperative effect on the recovery of the platelet after PBSCT. There is little side effect, and it can be accepted.

Based on the structural characteristics of Cycas trunk, including vegetative leaf base scars, sporophyll concave rings, and average occurrence probabilities of vegetative leaf and sporophyll, a method for the age estimation of Cycas fairylakea population was developed, and the age of each individual was calculated. Three approaches, i.e., age structure diagram, age distribution curve and curve estimation were used to study the age structure of C. fairylakea population at genet and clone population levels. The age structure diagram showed that the clone population of C. fairylakea was stable, but the genet population was in declining. However, both of the clone and genet populations were in declining when using the other two approaches. It was considered that the C. fairylakea population was in declining, and needed an urgent protection.

Objective: To determine the effect of different doses of atorvastatin on the serum soluble intercellular adhesion molecules-1 (sICAM-1) in patients undergoing percutaneous coronary intervention (PCI)., Methods: The study consisted of 38 patients with unstable angina and 10 patients with old infarction who underwent elected PCI for stenotic lesions of the coronary artery. Patients were randomly assigned to either aggressive group or conventional one. After PCI the patients took atorvastatin 20 mg per day or 10 mg per day. Blood lipid profile was examined before, and 3 months after the PCI. SICAM-1 was examined before the PCI, 48 hours and 3 months after the PCI., Results: The total cholesterol and LDL-Cholesterol 3 months after the PCI in the 2 groups were lower than those before the PCI (P<0.01). The aggressive group showed greater reduction in concentrations of TC and LDL-C than the conventional group (P<0.01). The changes in concentrations of HDL-C between pre-PCI and 3 months after the PCI and TG were not obvious (P>0.05). sICAM-1 in the 2 groups 48 hours after the PCI significantly higher than that before the PCI (P<0.01). But sICAM-1 in the 2 groups 3 months after the PCI significantly lower than that before the PCI (P<0.01 or P<0.05). The aggressive group showed greater reduction than the conventional group (P<0.01). TC and LDL-C were positively correlated with sICAM-1(r=0.2413, r=0.2691, all P<0.05)., Conclusion: Atorvastatin 20 mg per day reduces TC, LDL-C, and sICAM-1 to a greater extent than atorvastatin 10 mg per day. The effect on sICAM-1 is partly related to reduce lipid profile.

Objective: To induce hematopoietic progenitor/stem cells of umbilical cord blood to differentiate into mature megakaryocytes and platelets in vitro and to investigate the mechanism of production of platelets., Methods: The CD34+ cells were sorted from umbilical cord blood by magnetic activated cell sorting (MACS) and then cultured in vitro with optimized medium to be differentiated into mature megakaryocytes and platelets. The cultured cells and the platelet-like particles were isolated from the culture and were checked by the fluorescence-activated cell sorter (FACS), immunohistochemistry assays, light microscope,electron microscope and platelet aggregation tests., Results: The cultured megakaryocytes were detected with proplatelets and both the cultured cells and the platelet-sized particles were found to have the same structure with the normal megakaryocytes and platelets by light and electron microscope. The immunohistochemistry assays revealed the cultured cells expressed GP II b III a with a positivity of 95% which was a special antigen for platelets and megakaryocytes. Culture-derived platelet-sized particles aggregated in response to thrombin as the plasma derived-platelets did. The cultured platelets had the same positivity of CD41 as the platelets from platelet rich plasma., Conclusion: The hematopoietic progenitor/stem cells can be induced to differentiate into purified and mature megakaryocytes and platelets. It provides a practical way to study the mechanism of platelets production.

The aim of this study was to sort the CD34(+)/CD123(+) cells from the bone marrow cells of patients with acute myeloid leukemia (AML) by Midi MACS method. Firstly, the bone marrow mononuclear cells (BMMNC) were isolated from the patients with AML with Ficoll Paque, CD34(+) cells were then isolated by Midi MACS method followed by the isolation of CD34(+)/CD123(+) cells from the fraction of CD34(+) cells. The enrichment and recovery of CD34(+) and CD34(+)/CD123(+) cells were assayed by FACS technique. The results showed that the enrichment of CD34(+) cells was up to 98.73%, its average enrichment was 95.6%, and the recovery of CD34(+) was 84.6%, its average recovery was 51% after the first round sorting, by the second round sorting, the enrichment of CD34(+)/CD123(+) cells was up to 99.23%, its average enrichment was 83%. With regard to BMMNCs before sorting, the recovery of CD34(+)/CD123(+) was 34%. But, on the CD34(+) cells obtained by the first round sorting, its recovery was 56%. In conclusion, these results confirmed that the method of Midi MACS sorting can be applied to sort CD34(+)/CD123(+) cells from the bone marrow cells of AML patients, which give rise to the similar enrichment and recovery of the sorted cells with that of literature reported by the method of FACS.

This study was aimed to explore the possibility of rAAV2 vector-mediating gene therapy for Glanzmann' s thrombasthenia. The rAAV2-GPIIb/IIIa vector was constructed. The GPIIb/IIIa gene expression in mammal cell were examined by different methods, such as: detection of mRNA expression in BHK-21 cells after 24 hours of infection (MOI = 1 x 10(5) v.g/cell) was performed by RT-PCR; the relation between MOI and quantity of GPII6/IIIa gene expression was detected by FACS after 48 hours of infection; GPIIb/IIIa protein expression in BHK-21 cells after 48 hours of infection (MOI = 10(5) v x g/cell) was assayed by Western blot, GPIIb/IIIa protein expression on cell surface was detected by immunofluorescence, and the biological function of expressing product was determined by PAC-1 conjunct experiments. The results showed that GPIIb/IIIa gene expression in mRNA level could be detected in BHK-21 cells after 24 hours of infection at MOI = 1 x 10(5) v x g/cell and the GPIIb/IIIa gene expression in protein level could be detected in BHK-21 cells after 48 hours of infection at MOI = 1 x 10(5) v x g/cell. In certain range, quantity of GPIIb/IIIa gene expression increased with MOI, but overdose of MOI decreased quantity of GPIIb/IIIa gene expression. Activated product of GPIIb/IIIa gene expression could combined with PAC-I, and possesed normal biological function. In conclusion, rAAV2 vactor can effectively mediate GPIIb and GPIIIa gene expressing in mammal cells, and the products of these genes exhibit biological function. This result may provide a basis for application of rAAV2 vector in Glanzmann's thrombasthenia gene therapy in furture.

Objective: To investigate the expression of human coagulation factor IX (hFIX) gene in human umbilical cord blood (CB) CD34+ cells which was transfected with recombinant adeno-associated virus 2 (rAAV-2)., Method: The CD34+ cells were transfected with rAAV-2/hFIX and cultured for 21 days for inducing differentiation into myeloid, erythroid and megakaryocytes, respectively. The expression of hFIX was studied at mRNA, protein and biological activity levels. The cytotoxicity of rAAV-2 to CD34+ cells was evaluated by cell proliferation, cell vitality, CD antigen expressions and CFU yields., Results: The hFIX mRNA was expressed in the cultured cells which was verified by RT-PCR and DNA sequencing. An elevated level of hFIX expression and biological activities were detected with a maximum amount of 14.10 ng/10(6) cell x 24 h. During the period of 21 day culture, the cell vitality, cell proliferation, CD antigen expression and CFU yields between the transfected and un-transfected groups had no difference(P > 0.05)., Conclusion: The human CB CD34+ cells are able to produce functional hFIX after transduction by rAAV-2/hFIX. The cell proliferation and differentiation capacities of the host CD34+ cells were not affected by rAAV-2.

Objective: To explore whether normal platelet contains tissue factor (TF), and the significance of platelet-associated TF (PATF)., Methods: Platelets were isolated by Sepharose 2B gel column. ELISA was used to detect the TF content in the lysates of washed platelets. Procoagulant activity of PATF was measured by one stage clotting time assay. The mRNA of TF was detected by reverse transcription polymerase chain reaction (RT-PCR)., Results: A certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in the washed-platelet lysates. Upon activation by collagen, platelets released TF and caused a marked increase in TF level in plasma (P <0.05). Resting platelets had no TF procoagulant activity, while procoagulant activity of platelets activated by collagen increased significantly, which could be blocked by TF McAb and poor VII plasma. TF mRNA could not be detected in washed platelets. TF content in platelets from patients with coronary heart disease was significantly higher than that from normal controls (P < 0.05). Resting platelets from the patients showed a higher procoagulant activity, which could be inhibited by TF McAb., Conclusion: Platelets contain TF and the latter released by activated platelet was functionally active. Platelet itself might not synthesize TF. Protein content and procoagulant activity of PATF in patients with coronary heart disease were higher than that in controls. All these indicate that platelet may be involved in coagulation and thrombosis by releasing TF.

Objective: To explore the change in the protein content and procoagulant activity of platelet-associated tissue factor (TF) in thrombotic diseases., Methods: Platelets were isolated with Sepharose 2B gel column. ELISA was employed to detect the TF protein content in the lysates of washed platelets, procoagulant activity of platelet-associated TF was measured with one stage clotting time assay., Results: A certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in resting platelets in normal controls. The procoagulant activity of resting platelets and platelets activated by collagen were (0.89 +/- 0.26), (2.27 +/- 0.24) U/ml respectively. While specific TF McAb was added, the procoagulant activity of activated platelets can be markedly inhibited (1.39 +/- 0.28) U/ml (P < 0.001). It is thus proved that the procoagulant activity of activated platelets mainly comes from TF. TF protein content of the lysates of washed platelets in coronary heart disease group, brain infarction group and diabetes mellitus group was (20.71 +/- 8.78), (23.83 +/- 8.03), (22.12 +/- 8.24) ng/L respectively. The procoagulant activity of platelets without collagen treatment in these thrombotic diseases was (1.71 +/- 0.24), (1.69 +/- 0.25), (1.75 +/- 0.31) U/ml respectively. When specific TF McAb was added specific TF McAb, the procoagulant activity was (1.43 +/- 0.25), (1.39 +/- 0.25), (1.40 +/- 0.29) U/ml respectively. The TF protein content of lysates of washed platelets and the procoagulant activity of platelets without collagen treatment in patients with thrombotic diseases were significantly higher than those in normal controls. This procoagulant activity can be inhibited by TF McAb (P < 0.01)., Conclusion: The change in platelet-associated TF in thrombotic diseases may contribute to the maintenance of hypercoagulability.

The nuclear factor kappa B(NF-kappaB) plays a crucial role in inflammatory, immune response, embryo development, cell proliferation and apoptosis, cell cycle control as well as tumorgenesis. In recent years, a variety of investigations have demonstrated that NF-kappaB was closely associated with the pathogenesis of hematological malignancies such as leukemia, lymphoma and multiple myeloma. Nowadays, increasingly attention has been paid to the studies on the activation and its mechanism of NF-kappaB in the hematogenic malignancies. So that, in this article, progress on these aspects is reviewed.

To explore the effect of glycosyl-phosphatidyl inositol-specific phospholipase D (GPI-PLD) on the adhesion function of bone marrow mononuclear cell from patients with myeloid leukemia and analyze its mechanism, the activity of GPI-PLD in bone marrow mononuclear cell from the patients were measured by using GPI-anchored placental alkaline phosphatase (PLAP) as substrate and Triton-X114 partitioning; the adhesion rate and CD24 expression of these cells were measured by MTT and immunohistochemical method respectively, when these cells were or were not treated by 1 mmol/L 1,10-phenanthroline for 5 hours. The results showed that the GPI-PLD activity of bone marrow mononuclear cells from the patients was significantly inhibited after being treated by 1 mmol/L 1, 10-phenanthroline for 5 hours [(42.08 +/- 7.21)% vs (5.4 +/- 2.96)%], while the adhesion rate and the expression of CD24 of these cells were increased [(49.78 +/- 26.73)% vs (61.19 +/- 29.14)%, (16.02 +/- 9.68)% vs (18.5 +/- 11.14)%, respectively)]. It is concluded that depression of GPI-PLD activity can increase the adhesion rate of bone marrow mononuclear cells from the patients while the CD24 expression is enhanced.

To investigate the effect of GFP fused to C terminal of integrin alpha(IIb) on the biosynthesis and expression of alpha(IIb) beta(3) compound, the alpha(IIb) GFP expression plamid, named palpha(IIb) GFP, the cDNA of alpha(IIb) was constructed from p3.1-2b and fused to pEGFP-N1 in frame. When the sequence of palpha(IIb) GFP was confirmed by sequencing it was transferred to Chinese Hamster Ovary (CHO) cells with or without p3.1-3a expressing integrin beta(3). Then the expression of alpha(IIb) GFP fusion protein was confirmed by Western blot and then its subcellular localization was determined with laser confocal scanning microscopy. The results showed that the target gene was cloned into recombinant vector by restriction analysis and sequencing. Overexpression of the fusion protein in the transfected CHO cells was identified with Western blot. Subcellular localization analysis confirmed that alpha(IIb) GFP was expressed in CHO cells and could be transferred from endoplasmic reticulum to Golgi apparatus. It is concluded that the eukaryotic expression plasmid containing alpha(IIb) GFP fusion gene is successfully constructed. GFP fused to the cytoplasmic tail of integrin alpha(IIb) allows the normal expression of alpha(IIb) beta(3) in CHO cells.

Objective: To study the effect of glycoprotein (GP) alpha II bA2334C mutation on the biosynthesis and expression of alpha II bbeta3 complex., Methods: The GP alpha II bA2334C eukaryotic expression plasmid pc3.1-2334M2b was constructed. Chinese hamster ovary (CHO) cells were transfected with the plasmid with or without integrin beta3 expression plasmid pc3.1-3a. The whole expression of alpha II bA2334C was confirmed by Western blot and the membrane expression was analyzed by flow cytometry. A newly constructed alpha II bA2334C GFP fusion protein expressing plasmid was used to determine its subcellular localization by laser confocal scanning microscopy., Results: Expression of the mutant protein, alpha II bA2334C, in the transfected CHO cells was confirmed by Western blot with a lower rate of the mature type than the wild type control. The expression on membrane was only 25% of the normal. Subcellular localization analysis showed that alpha II bA2334C GFP was able to be expressed in CHO cells and could be transported from endoplasmic reticulum to Golgi apparatus., Conclusions: The mutant alpha II bA2334C can be synthesized in CHO cells and form alpha II bbeta3 complex. However, only a small fraction of the premature alpha II bA2334C can be transported to Golgi apparatus and transformed to mature alpha II b. The possible pathogenesis of this type II thrombasthenia may be that the misfolded alpha II bA2334C is partially degraded in the endoplasmic reticulum causing lower expression of alpha II bbeta3 complex on the membrane and resulting in impared function of platelets than normal alpha II b.

Objective: To prepare the rAAV2/hFIX and evaluate the efficiency of the preparation on gene therapy of hemophilia B model mice., Methods: The rAAV-2/hFIX was prepared by "one helper virus-one vector cell line" strategy and transfected both BHK-21 and C2C12 cells in vitro. The hFIX antigen level in cell culture supernatant was assayed. The rAAV-2/hFIX was injected into muscles of hemophilia B model mice and assayed the serum hFIX levels, hFIX clotting activity, bleeding time, 5 min bleeding volume., Results: The hFIX antigen could be detected from 24 h till 120 h after BHK-21 and C2C12 cells were transfected with highest levels at 24 h reaching (51.0 +/- 6.5) ng/10(5) cells and (68.0 +/- 7.2) ng/10(5) cells, respectively. The rAAV2/hFIX injected mice could efficiently express hFIX and peaked at three weeks after injection, then slowly decreased but low level hFIX antigen was still detectable till 10 weeks after injection. There were significant differences between the high, middle and low dose groups of rAAV2/hFIX and the control group (P < 0.01), the plasma FIX clotting activities in the model mice were improved remarkably, bleeding time was greatly shortened and bleeding in 5 min was decreased. The hFIX expression level and FIX clotting activity of the high dose of rAAV2/hFIX group (1.6 x 10(13) v.g./kg) reached about (387.0 +/- 12.5) ng/ml plasma in contrast with the normal levels of (30.0 +/- 5.5)% at the third week after injection. No rAAV2 vector DNA was detected in the organs except for injected muscle tissue., Conclusion: The rAAV2/hFIX transfected BHK-21 and C2C12 cells could efficiently express hFIX antigen and was of therapeutic effects for the hemophilia B model mice by intramuscularly injection.The results provide the basis for clinical trial of rAAV2 gene therapy for hemophilia B.

The aim of this study was to apply a new type piezo-electric immunosensor in immunology classification for acute leukemia. Based on the plasma-polymerized film and the nanogold particles self-assembly technology, a new type piezo-electric immunosensor was firstly developed to absorb and fix the monoclonal antibodies for detecting leukemia cell antigens in patient blood samples. The results showed that the positive rate of detection with this method was quite close to that with the fluoroimmunoassay which are commonly used in clinic work. There was no significant difference between the positive rates (chi(2) = 3.4, P > 0.05). It is concluded that this piezo-electric immunosensor detection has many advantages, such as convenient operation, real time operation and control, qualitative detection, and less cost. This method can be used for acute leukemia immunology classification.

Objective: To explore the dynamic change of Phlegm-stasis in the rat atherosclerotic model as the time goes on., Methods: Adopting high fat forage fed to develop the atherosclerotic model in rats, and the changes of blood lipid, hemorrheology, blood glucose, insulin and vascular smooth muscle cell (VSMC) actin expression were detected by biochemical and immunohistochemical assay at various time points after modeling., Results: The expression of VSMC actin gradually increased along with the change of model rats' Syndrome from Phlegm to stasis, i.e., the change of parameters, including blood lipid, hemorrheologic parameters, blood glucose, insulin and insulin sensitive indexes along with the aggravation of disease., Conclusion: The expression of VSMC actin could be the molecular mechanism for the Syndrome developing from Phlegm to stasis in atherosclerotic rats.

To investigate the transduction efficiency of recombinant adeno-associated virus 2 (rAAV-2) in human umbilical cord blood CD34(+) hematopoietic stem/progenitor cells, the CD34(+) cells sorted by the method of magnetic cell sorting from human cord blood were infected with the rAAV-2 expressing the green fluorescent protein (GFP) gene. After transduction for 19 hours, the expression of GFP was detected under fluorescence microscope. The results showed that 43% CD34(+) cells expressed the GFP gene at a multiplicity of infection of 2 x 10(5). It is concluded that the rAAV-2 can transduce human cord blood CD34(+) hematopoietic stem/progenitor cells efficiently.

Objective: To establish the possible relationship between some coagulation factors and the onset of acute cerebral infarction (ACI)., Methods: The study population consisted of 71 patients with ACI confirmed by CT and 50 age-matched healthy volunteers. Blood samples were obtained during the onset period of ACI. Tissue factor (TF) and tissue factor pathway inhibitor (TFPI) activity in plasma were assayed with the chromogenic assay. Plasma TF and TFPI antigen were measured with enzyme linked immunoadsorbent assay (ELISA). Plasma F VII coagulation activity (F VII: C) and F VIII coagulation activity (F VIII: C) were developed in the one-stage system. Plasma prothrombin (FII) was determined with Ecarin assay. Plasma fibrinogen (Fbg) was measured with thrombin assay. Plasma antithrombin III activity (ATIII) was determined using heparin cofactor activity assay., Results: Compared with the control, plasma TF activity and antigen in patients with ACI were significantly higher (both P<0.05). But plasma TFPI activity and antigen were remarkably lower in the ACI group (both P<0.05). Plasma F VII: C was significantly higher (P<0.01), and F VIII: C was markedly lower (P<0.05). Plasma FII was remarkably higher (P<0.01). Similarly the Fbg was significantly higher in the ACI than that in the control group (P<0.01), whereas ATIII was significantly lower (P<0.01)., Conclusion: The initiation of TF pathway is contributed to the onset of ACI and the blood is in hypercoagulable state during the early period of ACI.

Objective: To elucidate the effect of angiotensin II (AngII) on the expression of tissue factor (TF) by monocytes and its mechanisms., Methods: Monocytes were isolated from healthy volunteers by Ficoll-Hypaque gradient and Percoll, and cultured in RPMI-1640. Procoagulant activity (PCA) was determined by one-stage clotting method, TF antigen by ELISA. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the TF gene mRNA. The levels of IkappaBalpha was detected by Western blot. Electrophoretic mobility shift assays (EMSA) were performed to evaluate the activity of NF-kappaB., Results: AngII (10(-9) - 10(-7) mol/L) significantly increased monocyte PCA, TF antigen and TF mRNA expression in a dose and time dependent manner. Losartan (10(-6) - 10(-5) mol/L) significantly inhibited the effects of AngII on TF activity, antigen and mRNA expression in a dose-dependent effects. Staurosporine (2.5 x 10(-7) mol/L) and genistein (4 x 10(-5) mol/L) lowered TF level of monocytes (P < 0.05). Western blot analysis revealed that after exposure to AngII (10(-7) mol/L), IkappaBalpha level decreased at 15 min, reached nadir at 60 min, and recovered at 180 min. EMSA showed NF-kappaB binding activity increased at 15 min, reached peak at 60 min, and recovered at 180 min. Pyrrolidine dithiocarbamate (PDTC, 10(-4) mol/L), an inhibitor of NF-kappaB, or AT1R antagonist losartan (10(-5)mol/L) inhibited AngII-induced NF-kappaB translocation., Conclusions: AngII could induce the expression of TF in human monocytes, and this effect was mediated by AT1R. The PKC pathway played the most important role in AngII-induced TF expression. The activation of NF-kappaB was involved in TF expression in monocytes.

To study the relationship between the level of the soluble L-selectin (sL-selectin) in plasma and surface L-selectin expression on leukemic cells and episode and state of illness in acute leukemia patients, the plasma level of sL-selectin was measured by a sandwich enzyme-linked immunosorbent assay, and the expressions of surface L-selectin and its gene (lyam-1) were detected by immunohistochemistry and RT-PCR. The results showed that the levels of sL-selectin were significantly higher in untreated and therapy-resistant acute leukemia patients, and expression of L-selectin mRNA and cell surface L-selectinin in untreated and NR patients were significantly lower than that in CR patients and control group (P < 0.05). The plasma levels of sL-selectin were significantly increased in patients with splenomegaly and hepatomegaly (extramedullary infiltration). The levels of sL-selectin were related to the clinical course of the acute leukemia patients. A significant correalation existed between expressions of L-selectin mRNA and surface L-selectin in acute leukemia patients (gamma = 0.782, P < 0.05). It is concluded that expression of L-selectin gene was down-regulated in level of mRNA and protein in acute leukemia patients and both changes were highly correlated. Monitoring of the plasma level of sL-selectin is possibly useful for early diagnosis of relapse and extramedullary infiltration in acute leukemia.

Objective: To study the signal transduction in the course of energy metabolism of platelets, and to provide a new theoretical basis for clinical application of the anti-activation of platelets by the monoantibody of integrin GPIIb/IIIa., Methods: With or without the block of integrin GPIIb/IIIa monoantibody 10E5 and/or integrin alpha v beta 3 monoantibody 609, the uptake rate of tritium-labeled 2-deoxyglucose([3H]2-DG) by platelets activated by thrombin was counted in a liquid scintillation counter. The relationship between the energy metabolism of platelets and platelet integrin was investigated by the course of glucose uptake of individual vivo platelets., Results: Compared with unactivated platelets, the glucose uptake of activated platelets increased, and it could be blocked by monoantibody 10E5 and partly blocked by 609., Conclusion: The glucose uptake of activated platelets can be suppressed by monoantibody 10E5, and the intergrin family is associated with the signal transduction of platelet energy metabolism. Integrin GPIIb/IIIa may play an important role in the procedure of platelet energy metabolism.

Objective: To compare the different effects of the programmable cell freezer and the -70 degrees C mechanical freezer in the preservation of the combilical cord blood and to study the effects of magnetic activated cell sorting on cell biologic characters., Methods: After separating the nucleated cells from the umbilical blood by the density gradient separation method, we used cryoprotectant to protect the nucleated cells and preserve them by two methods; one was using the programmable cell freezer to cool the cells and finally putting them to the liquid nitrogen, and the other one was directly preserving the cells in a -70 degrees C mechanical freezer. The cells were thawed after a month and tested for the viability and clonogenic ability. Cell recovery, clonogenic capacity and clonogenic recovery by the two cryopreservation methods were compared. Meanwhile, the CD34+ cells were sorted by the method of magnetic cell sorting from the umbilical cord blood, and the cell purity and recovery after sorting were analyzed by the method of fluorescence-activated cell sorting., Results: There were no significant differences between the two preserving methods in cell viability, cell recovery, clonogenic capacity, or colony recovery. The purity of the CD34+ cells was 88.3% and the recovery rate was 46.2% after the cell sorting. The cell viability and clonogenic capacity of the CD34+ cells were much higher than those of the CD34- cells., Conclusion: The umbilical cord blood can be efficiently preserved by the two methods and highly purified CD34+ cells could be obtained by using the method of magnetic cell sorting. The procedure of cell sorting doesn't affect the viability and clonogenic ability. The study provides some evidence for clinical application of preserving the umbilical cord blood and transplanting CD34+ cells.

Objective: To evaluate the efficacy of hematopoietic stem cell transplantation (HSCT) in hematological malignancies., Methods: Twenty-five patients with hematological malignancies were treated by autologous bone marrow transplantation (ABMT), autologous peripheral blood stem cell transplantation (APB-SCT) or Allo-PBSCT. Eight acute myeloid leuekmia (AML) patients who received autologous hematopoietic stem cell transplantation (AHSCT) were treated by chemotherapy again., Results: All the patients engrafted gained hematopoietic reconstitution; they were followed up for 6 to 91 months; five patients died; twenty patients were alive in a disease-free situation., Conclusion: After the patients with acute leukemia receive AHSCT, the relapse incidences can be reduced obviously by regular chemotherapy. HSCT is one of the best methods for hematological malignancies.

Objective: To investigate the effect of the defect of integrin alpha II b beta 3 on the inside-out signal transduction in platelets., Methods: The transfected cDNA, its expression and the ability of cells binding to PAC-1 and fibrinogen were investigated by RT-PCR, DNA sequence analysis, flow cytometry and Western blotting., Results: The integrin alpha II b beta 3 level in the patients with Glanzmann's thrombasthenia was significantly lower than that of the normal subjects and the platelets of the patients failed to bind PAC-1 activated by ADP. There were both C3077G and G3078C mutations in exon 30 of mutant alpha II bR995A beta 3 cDNA, which resulted in an amino acid substitution arginine (R) 995 to alanine (A). CHO cells transfected with wild-type alpha II b beta 3 or mutant alpha II bR995A beta 3 cDNA respectively expressed normal alpha II b beta 3 and mutant alpha II bR995A beta 3. When cells were not activated, wild-type alpha II b beta 3 CHO cells failed to bind PAC-1, but could adhere to fibrinogen, but mutant chimera alpha II bR995A beta 3 CHO cells were able to bind PAC-1 and exhibited high affinity binding fibrinogen., Conclusion: The point mutation in integrin alpha II bR995A can induce the inside-out signal transduction in platelets, and have integrin alpha II bR995A beta 3 locked into an activation state; the defect of alpha II b beta 3 in patients with GT impairs the inside-out signal transduction mediated by alpha II b beta 3 in platelets and the adhesive functions of platelets.

Objective: To determine the occurrence of activated protein C resistance (APCR), to identify APCR is associated with thrombotic events (TEs), and acquired APCR is associated with the presence of antiphospholipid antibodies (APLAs) in 30 patients with systemic lupus erythematosus (SLE)., Methods: Laboratory tests included dilute Russell's viper venom time assay for LA (dRVVT-LA), ELISA assay for ACL, APC sensitivity ratio, and factor V Leiden were detected by PCR-Mnl/I digestion., Results: Acquired APCR was presented in 14(46.67%) of 30 patients. Factor V Leiden was not found in any patients. The incidence of TEs in the APCR-positive patients was significantly higher than that in the APCR-negative patients (42.85% vs 6.25%, P < 0.05). The incidence of TEs in the LA-positive patients was also significantly higher than that in the LA-negative patients (50% vs 11.1%, P < 0.05)., Conclusions: The presence of either APCR or LAs is associated with one of the risk factors of TEs (P < 0.05). There is not a significant interaction between APCR and LAs in the association with TEs. Acquired APCR may not reflect the interference of LAs with the protein C pathway which may represent a mechanism of LA-associated TEs.