A prospective study of the efficacy and safety of maintenance therapy with recombinant human thrombopoietin in patients with primary immune thrombocytopenia: a multicenter study / 中华血液学杂志

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≥50×109/L were enrolled to maintenance therapy starting with every other day administration of rhTPO, then adjusted dose interval to maintain platelet count (30-100) ×109/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≥30×109/L for 12 weeks. The percentage of patients with a platelet response (PLT≥30×109/L) at 4th week, 8th week and 12th 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) ×109/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.

Observation of the effed of tiotropium bromide combined with ambroxol in of the treatment of chronic ob-structive pulmonary disease / 中国基层医药

Objective To explore the effect of tiotropium bromide combined with ambroxol hydrochloride oral liquid on chronic obstructive pulmonary disease (COPD) at remission.Methods According to the digital table, 38 patients with COPD at remission were randomly divided into treatment group and control group .The control group was given with conventional treatment ,and anti-infection and symptomatic treatment when acute attacking ,while the treatment group was treated with inhaled tiotropium bromide and ambroxol Hydrochloride oral liquid base on the con -trol group.Both groups were treated and observed for 6 months.Times of expectoration and acute attack of COPD were observed,and scores of COPD Assessment Test ( CAT) were noted everyday during therapy .Results Compared with the control group ,the total times and means of expectoration in treatment group were much less ,with very significant statistical difference(t=5.19,P<0.05);the cases,times and means of acute attack of COPD were obviously less than those in control group,with very significant statistical difference (t=2.92,P<0.05);the total scores and means of CAT in treatment group were much lower,with significant statistical difference(t=2.65,P<0.05).Conclusion With long-term treatment ,inhaled tiotropium bromide combined with ambroxol hydrochloride oral liquid can effectively decrease the times of expectoration and acute attack in patients with COPD at remission,and improve their qualities of life.

Real-time quantitative PCR monitoring of minimal residual disease in CML after treatment / 白血病·淋巴瘤

Objective To set up a real-time quantitative PCR approach for detection and quantification for bcr-abl transcripts in CML patients,and detect minimal residual disease (MRD) in CML by real-time quantitative PCR (RQ-PCR)and evaluate the significance of MRD detection.Methods The ber-abl.fusion gene expression in 80 patients with CML was analyzed by RQ-PCR. The patients were divided into three groups according to the different treatment, allogeneic hematopoietic stem cell transplantation group,imatinib group and hydroxyurea group. The change of bcr-abl fusion gene was monitored in CML patients before and after treatment.Results The average of RQ-PCR detection on newly diagnosed patients with CML in chronic phase was 6847.67 copies / 104 cells,the accelerated phase was 306 176.08 copies / 104 cells,and the average results were 944.33, 2.37, 0.29, 0 copies / 104 cells after allogeneic hematopoietic stem cell transplantation one month,6 months,12 months or 24 months respectively.The average of RQ-PCR detection after use imatinib mesylate 3 months was 3720.23 copies / 104 cells and not be detected after one year. The average was 7290.11 and 3143.24 copies / 104 cells after hydroxyurea treatment 0 and 9 months respectively.The difference in first two groups was not significant (t=1.74,P=0.17), but the difference between the third group and the first two groups was significant (t=3.74,P=0.01.t=2.97,P=0.02). The upregulation of bcr-abl transcript levels could be detected when disease progression. The transcripts level in accelerated phase was significantly higher than that in chronic phase. Conclusion RQ-PCR can be used to detect the MRD,monitor the treatment outcome,predict disease recurrence and give early intervention.

Study on chromosomal aberrations in patients with multiple myeloma by fluorescence in situhybridization / 白血病·淋巴瘤

ObjectiveTo explore the genetic abnormalities of multiple myeloma (MM)patients by fluorescence in situ hybridization (FISH).MethodsWith the application of FISH,sequence specific DNA probes (IGH,DI3S319/p53 and 1q21/RB1) were applied to detect 14q32 rearrangement,del(13q14),del (17p13)and gain of lq21.Forty-four MM patients were enrolled in this study.ResultsThirty-two cases (72.7 ％) detected by FISH had genetic abnormality in 44 cases,lq21 amplification was observed in 11 cases (25.0 ％),while RB1 deletion in 17 cases (38.6 ％),D13S319 deletion in 16 cases (36.4 ％),p53 deletion in 6 cases(13.6 ％)and 14q32 translocation in 19 cases(43.2 ％).The patients with one abnormality was detected in 10 cases(22.7 ％),two abnormalities in 11 cases(25.0 ％),three abnormalities in 8 cases (18.2 ％),4 abnormalities in 3 cases(6.8 ％).28 were found to undergo split-phase by conventional cytogenetic in 44 patients.The patients with genetic abnormalities detected by conventional G-banding was 2 cases (7.14 ％),the difference with that in FISH was significant (P ＜0.05).Genetic abnormalities compared with clinical parameters showed that β2-MG in IGH gene abnormal patients were significantly higher than those without such abnormalities (P ＜0.05).Patients with bone marrow plasma cells of lq21 amplification were higher than those with normal karotypes(P ＜0.05),CRE was significantly higher among lq21 amplification and p53 deletion patients (P ＜0.05),CRP was significantly higher among p53 deletion patients (P ＜0.05).No significant difference was oberserved in relationship of the chromosome aberration and age,the chromosome aberration and stage.ConclusionThe most common genetic abnormalities in MM is IGH rearrangement and absence of RB1 and D13S319,followed by lq21 amplification,the least is p53 deletion.FISH is a rapid and sensitive technique to refine chromosome aberrations in MM.The specific detection for genomic features of MM is proved to be correlative with its clinicopathologic characteristics and the prognosis.

Clinical study on modified GMALL protocol for adult acute lymphoblastic leukemia / 白血病·淋巴瘤

Objective To analyze the treatment efficacy and safety of a modified GMALL protocol for adult acute lymphoblastic leukemia (ALL). Methods Data of 37 patients with newly diagnosed adult ALL treated with a modified GMALL protocol from January 2005 to December 2009 were retrospectively analyzed,and compared with that of 44 patients treated with an in-house conventional protocol at the same period.Results The complete remission (CR) rate was 89.2 %(33/37) treated with modified GMALL protocol. The cumulative overall survival (OS) rates at 1 year, 2 years, 3 years and 4 years were 77.5 %, 48.0 %, 40.0 %and 40.0 %, respectively. The main adverse events were grade 3 or grade 4 hematological toxicities and infections which were easily managed, and the treatment-related mortality rate was low. The OS of modified GMALL protocol was superior to that of the conventional protocol. Conclusion The modified GMALL protocol has a satisfying effect and the adverse events can be tolerated for adult ALL, so its clinical application can be encouraged.

Clinical analysis of mixed acute leukemia / 白血病·淋巴瘤

Objective To analyze the clinical and biological features of mixed acute leukemia(MAL).Methods Bone marrow specimens of 38 MAL patients were evaluated to prove the diagnosis and the classification by morphoiogic,immunologic examinations.These patients were treated with protocols suitable for both acute myeloid leukemia(AML)and acute lymphoblastic leukemia(ALL).Results All MAL patients had a leukemia syndrome.Morphologically,the subtypes of M1,M2 and M5 were predominant in AML,as L2 Was in ALL.Immunologically,coexpression of myeloid and B lineage associated antigens was predominant,about 68.4%;cytogenetically,Ph chromosome was observed in 33.3%(5/15)of MAL patients,and immunophenotype was B-M;1 Ph chromosome(+)MAL patient,fusion gene bcr-abl 190(+)and immunophenotype was B-M.In 38 cases,32 patients received chemotherapy.The complete remission rate was 28.1%(9/32).CR of.normal karyotype was significantly higher than that of abnormal ones.Conclusion Patients with MAL have unique biological features and the complete remission rate was low and the prognosis was poor.

Treatment of two case childhood acute lymphoblastic leukemia by HLA-mismatched unrelated umbilical cord blood transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the hematopoietic and immunologic reconstitution and transplantation-related complications of HLA one locus mismatched unrelated umbilical cord blood transplantation for the treatment of hematological malignancies.</p><p><b>METHODS</b>Two children with acute lymphoblastic leukemia received HLA-mismatched unrelated umbilical cord blood transplantation. The conditioning regimens were BU-CTX (case 1) and BU-CTX plus BCNU (case 2). GVHD prophylaxis regimen consisted of cyclosporine (CsA) and mycophenolate mofetil (MMF). The patients received 14.6 x 10(7) nucleated cells/kg with 7.24 x 10(5) CD(34)(+) cells/kg and 16.24 x 10(7) nucleated cells/kg with 21.11 x 10(5) CD(34)(+) cells/kg, respectively.</p><p><b>RESULTS</b>The two recipients, ANC > 0.5 x 10(9)/L occurred at day 27 and day 17, BPC > 50 x 10(9)/L at day 53 and day 46, the peripheral blood counts normalization at day 60 and day 52, the immune function normalization at day 134 and day 122 and the DNA fingerprinting showing engraftment at day 19 and day 17, respectively. The donor-recipient pair of case 1 was male to female, and the chromosome karyotype of recipients bone marrow and peripheral blood cells showed 100%, 46, XY cells at day 49. Grade II acute graft versus host disease (aGVHD) occurred at day 26 (case 1) and day 21 (case 2). The two recipients have survived for 353 days and 256 days.</p><p><b>CONCLUSION</b>The hematopoietic and immunologic reconstitution in umbilical cord blood transplantation were earlier and more durable. The transplantation-related complications were less and aGVHD were milder.</p>

Detection of platelet fibronectin from congenital fibrinogenopenic patients and its clinical significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To assess the platelet and plasma concentrations of fibronectin (Fn) and fibrinogen (Fg) in congenital fibrinogenopenic (FgP) patients and explore their role in inducing platelet adhesion and aggregation.</p><p><b>METHODS</b>A FgP family was selected as study group and the platelets isolated and purified to assess concentrations of Fn and Fg in platelets, alpha-granules and plasma with Western blotting, immuofluoresence staining and flow cytometry (FACS), respectively, the expression of platelets GP II b/III a by FACS.</p><p><b>RESULTS</b>The concentration of platelets Fn in FgP patients is higher than that in controls, and is higher in homozygote than in heterozygote. In contrast, plasma Fn levels were identical in all samples. The amount of platelet Fg from FgP patients is lower than that from the controls and positively correlated with the concentration of their plasma Fg. No difference in the expression of platelet GP II b/III a had been found.</p><p><b>CONCLUSION</b>It suggested that increased platelet Fn could partially compensate the lack of Fg and lead the platelet adhesion and aggregation.</p>

The relationship between anti-phospholipid antibodies and activated protein C resistance / 中华风湿病学杂志

Objective To investigate the relationship between the anti-phospholipid antibodies (APLs)and activated protein C resistance (APCR). Methods The response to activated protein C (APC) was studied by an APTT-based (clotting) assay with a Stago autoanalyzer and expressed as the ratio between the APTT obtained in the presence and absence of exogenous APC. APC sensitive ratio higher than 2 was regarded as APCR. Anti-?2-glycoprotein Ⅰ (?2GP-Ⅰ) antibody and anti-cardiolipin antibodies (ACL) were measured by an enzyme-linked immunosorbent assay (ELISA). Lupus anticoagulant (LA) was tested by activated partial thromboplastin time (APTT). Results The existence of LA and acquired APCR showed significant correlation (?2=16.332, P=0.008). Acquired APCR was significantly associated with the presence of anti-?2GP-Ⅰ antibody (?2=6.179, P=0.012), but not ACL. The presence of APCR was associated with an increased frequency of history of thromboembolic events and/or recurrent abortions (?2=7.347, P=0.01). Conclusion This study suggests that APCR is linked to the presence of LA and anti-?2GP-Ⅰ antibody. APLs may interfere with the activation of protein C. APCR phenotype may be a major risk factor for thrombophilia in patients with APLs. Combined detection of APLs has the potential value for predicting thrombosis.

Molecular cloning of human FL gene and its expression in E.coli / 中国生化药物杂志

Purpose　The aim is to obtain the cDNA sequence of encoding extramembrane human FL gene with high level expression in E.coli. Methods　The primers were designed based on the known FL cDNA sequence. The total RNA was isolated from fetal liver cells , and then RT-PCR was performed. The fragment was cloned into pUC-18T vector, and further sequenced by automatic sequence analyzer. The gene was inserted into GST fusion expression vector between BamH Ⅰ and EcoR Ⅰ sites. The recombinant plasmid was transformed into E.coli strain DH5 α and induced with 1mmol/L IPTG.Results　The 546bp DNA fragment was amplified by RT-PCR method from fetal liver cells and its sequence was identical to the published sequence encoding human FL. The expressed fusion protein, with molecular weight of about 22kD, was about 10% of the total bacteria protein by SDS-PAGE and densitometry analysis.Conclusion　cDNA was cloned successfully. This study provided a basis for the further fundamental research and clinical application of FL.

Study on Activated Protein C in Patients with Neuropsychiatric Lupus Erythematosus / 中华皮肤科杂志

Objective To explore the significance of activated protein C resistance (APCR) and antiphospholipid antibody(APA) in patients with neuropsychiatric lupus erythematosus(NPLE). Methods APCR, anticardiolipid antibody (ACA)(IgG, M, A), lupus anticoagulant (LA) were measured with APTT? APC, ELISA, PTT－ LA methods, respectively, in 21 NPLE patients and 88 SLE patients without NPLE(NNPLE). Results The positive rates of APCR and ACA(IgG) in NPLE group were 78.9％ (15/19) and 52.4％ (11/21), respectively, which were significantly higher than those in NNPLE group: 44.3％ (39/88) and 22.7％ (20/88), respectively (P