The mechanism of drug resistance about ATRA curing acute promyelocytic leukemia involves many aspects and was not understood completely now. Among them, gene, protein and cell signal conduction way have become hot points that scientists are focusing on recently.

ct curative effect and prognosis of multiple myeloma.

Objective To detect quantitatively hepatocyte growth factor(HGF)mRNA expressions of bone marrow mononuclear cells(MNCs)in acute leukemia(AL)and investigate its clinical significance.Methods Total mRNA of quantitated bone marrow MNCs isolated from 67 de novo AL cases was extrated and then cDNA was synthesized.Expression of HGF mRNA was quantified absolutely using real-time fluorescence quantification PCR(FQ-PCR).Results Expressions of HGF mRNA in a group of AL were higher significantiv than these in a control group(6.936 ±1.613,0.407 ±0.170,P<0.001),but there was similafitv between a group of acute myeloid leukemia(AMI,)and group of acute lymphoblastic leukemia (ALL)(7.127±1.911,6.635±0.934,P>0.05).In AL subtypes,the expression of M5(9.998 4±1.454)was higher than that of M2,M3,M4,L1,L2 and L3(P<0.001),but there ware no differences among the latters(P>0.05). Meanwhile,there was no statistical significance on the expressions of HGF mRNA between different age and sex(P>0.05).In addition,expressions of HGF mRNA in the remission group were lower than these in the non.remission group(6.393±1.165,8.041±1.848,P<0.005).Conclusions There are statistical significances of the expressions of bone marrow MNCs HGF mRNA among the AL group and control group.As to AL subtypes,there are no statistically significant differences between AML and ALL as well as between different age and sex.Besides,lower HGF mRNA level is correlated with better curative effect.It is suggested that HGF mRNA is a suitable index for AL diagnosis and treatment.

Objective To construct quantitative standard for quantification of hepatocyte growth factor (HGF) mRNA and establish its real-time fluorescence quantitative(FQ)-PCR assay to estimate its clinical relevance in lymphoma.Methods Recombinant plasmid Was constructed with target cDNA obtained from isolated total RNA by RT-PCR After PCR products were identified and purified,recombined plasmids were quantitated and then acted as quantitative standard.A new real time FQ-PCR analysis system Was established with the second pair of primers and the probe after amplification condition and the concentrations of components were optimized.HGF mRNA expressions in 47 lymohoma cases[11 Hodgkin disease(HD) cases,36 non-Hodgkin lyphoma(NHL)cases.among these patients,36 patients in remission while 11 patients without remission ] were analyzed quantitatively,and its specificity and sensitivity for lymphoma diagnosis were evaluated by receiptor operation character(ROC)curve method.Results HGF mRNA quantitative standard was constructed successfully.and its real time FO.PCR analysis system Was established combined with hot.start PCR and down.touch PCR technique. According to slope of standard curve (-3.513)and correlation cofficient(0.999),amplification efficiency of the system was 92.6%.Coefficient variation of intra-assay,intra-day and inter-day-assay were 2.1%,4.0% and 6.8%,respectively.Sensitivity of FQ-PCR Was 2 eopies/μl.Expressions of HGF mRNA in lymphoma group Was higher than that in control group(6.425±2.172 and 0.317±0.192,respectively,t=15.883,P<0.001),and its expressions in remission group was lower than no remission group(6.157±1.712 and 7.59l ±1.184,respectively,t=2.768,P<0.05).However,there Was not difference of HGF mRNA level between group HD and group NHL(P>0.05).According to ROC analysis,its sensitivity and specificity were 93.6% and 100% when cutoff value for lymphoma clinical diagnosis Was 3.136.Conclusion HGF mRNA'8 quantitative standard and its real time F9-PCR analysis system have been successfully constructed,and it can be used for quantitative detection of its mRNA expression in lymphoma.

AIM: To evaluate the expression of fibroblast growth factor receptor 3 (FGFR3) in acute lymphocytic leukemia (ALL) patients and its contribution to the proliferation of circulating endothelial cells (CECs).METHODS: The mRNA expression levels of FGFR3 in 44 patients with ALL were assayed by RT-PCR.Overall survival (OS) rates of the patients in FGFR3+ group and FGFR3-group were estimated by Kaplan-Meier analysis.The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cell counts, antigen expression, growth curve and colony forming rate of the CECs in the 2 groups were determined.The FGFR3 expression of CECs was identified by immunofluorescence staining.RESULTS: The positive rate of FGFR3 mRNA expression was 43.2% in 44 ALL patients with normal karyotype.T-ALL expressed higher level of FGFR3 than B-ALL (P<0.05).FGFR3 was over-expressed in ALL patients with bone marrow blast proportion ≥5% (P<0.05).The probability of OS was significantly lower in FGFR3+ group than that in FGFR3-group (P<0.05).The sorted CECs highly expressed CD31, CD144, VEGFR-2 and CD146, and rarely expressed CD45.The counts of CECs and expression level of CD133 significantly increased in FGFR3+ group compared with FGFR3-group.The same result of the amount of colony formation was observed (P<0.05).There was significant difference at 3 time points of cultured CECs count in vitro between FGFR3+ group and FGFR3-group (P<0.05).The positive rate of FGFR3 expression of CECs from 19 ALL-FGFR3+ patients was (29.00±15.71)%.CONCLUSION: The over-expression of FGFR3 gene in ALL may be helpful to evaluate the prolife-ration of CECs, and become a double target with anti-tumor and anti-angiogenesis effects to offer more choice for molecular therapy in the future.

Burkitt lymphoma (BL) is a highly aggressive B-cell non-Hodgkin lymphoma (NHL) derived from germinal center B cells, which can be divided into endemic, sporadic and immunodeficiency-associated types. BL is characterized by c-myc deregulation due to translocation of c-myc gene to an immunoglobulin enhancer region, resulting in excessive cell proliferation and rapid tumor progression, however the dysregulation of myc alone can't explain the pathogenesis and progression of BL. With the development of molecular biology technology in recent years, the further research on molecular mechanisms of BL pathogenesis may provide a theoretical basis of for clinical diagnosis and targeted drug therapy. This review discusses the progress of pathogenesis in BL.

Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38％ of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95％,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.

OBJECTIVETo delineate the clinical features and prognostic significance of acquired trisomy 21 (+21) in 31 patients with acute myeloid leukemia (AML).

METHODSChromosome specimen was prepared from bone marrow samples using a direct method and (or) cultivation, and their karyotypes were analyzed with R banding. The clinical features, chemotherapeutic effect and survival status of patients with acquired +21 were evaluated.

RESULTSCytogenetic studies were successfully performed on 3 329 patients with newly diagnosed AML, among which 31 (0.93%) had acquired +21. And 16 (0.48%) of the 31 patients had +21 as the sole abnormality. The most frequent subgroup of bone marrow morphology was AML-M5b, and its total number was 12 (38.7%) of the total. Thirty patients among those with +21 received standard chemotherapy. The complete remission (CR) rate (63% vs. 80%, P< 0.05) and median overall survival (OS) (7 months vs. 15 months, P< 0.01) of AML patients with acquired +21 were both lower than those without. Age (60 years or older) was associated with a significantly lower CR rate (30% vs. 80%, P< 0.05) and shorter median OS (4 months vs. 12 months, P< 0.01). Comparing to acquired +21 with other additional abnormalities, acquired +21 solely was associated with a lower median OS (6 months vs. 12 months, P< 0.05), but did not affect the CR rate (60% vs. 67%, P> 0.05). Three patients undergoing allogenetic hematopoietic stem cell transplantation (allo-HSCT) were still alive at the end of follow-up. Their survival time have reached 56, 36 and 105 months, respectively, which were remarkably longer than those only received chemotherapy (P< 0.01).

CONCLUSIONThe presence of acquired +21 in patients with AML has a adverse prognosis with or without other additional abnormalities. Older age (60 years or older) and +21 alone predicted relatively poorer outcome. Allo-HSCT was expected to prolong the survival time of AML patients with acquired +21.

Adolescent ; Adult ; Aged ; Chromosome Aberrations ; Down Syndrome ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Prognosis ; Young Adult

OBJECTIVETo explore the expression and clinical significance of Musashi2 (MSI2) gene in de novo acute myeloid leukemia (AML).

METHODSReal-time quantitative PCR (RQ-PCR) was used to measure the expression of MSI2 gene in 181 de novo AML patients. Correlation between the expression level and clinical features of such patients was explored.

RESULTSTranscript of the MSI2 gene was detected in 181 AML patients, with the median expression level being 2.341 (0.1124-58.8566). By contrast, CD34+ cells from 10 healthy controls had a much lower expression level (P=0.012), and the expression level of MSI2 in 24 patients with complete remission was significant lower than de novo patients (P=0.021). Based on the median expression level, such patients were divided into low expression group and high expression group. Patients from the high expression group had significantly higher rate of high white blood cell count (78% vs. 63%, P=0.034). Compared with MSI2-low group, FLT3-ITD mutation were much more common in MSI2-high group (28% vs. 7%, P=0.002). The expression level of MSI2 in aberrant karyotypes was much higher than that in favorable karyotypes (the median expression level was 2.7726 and 2.0733, P=0.035). Kaplan-Meier analysis showed that the overall survival in high expression group of MSI2 was lower than the low expression group, with the median survival time being 28 months and 12 months, respectively (P=0.045).

CONCLUSIONDe novo AML patients have a higher level of MSI2 gene expression. And the latter is much more common in those with high white blood cell count and aberrant karyotypes, and has a positive correlation with FLT3-ITD mutation. High expression of MSI2 gene may be a predictor for poorer prognosis among AML patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; RNA-Binding Proteins ; genetics ; metabolism ; Young Adult

Objective To observe the clinical effect of sequential therapy with micafungin and reduced -dose voriconazole in prevention of invasive fungal infections in patients received allogeneic hematopoietic stem cell transplantion (Allo -HSCT).Methods 28 patients received the treatments for prevention of fungal infection with micafungin 50 mg per day from pretreatment to 30 days,then oral voriconazole at a dose of 1 00 mg two times per day until 90 days after Allo -HSCT.The occurrence of invasive fungal infection and the side effects of both medicine were observed during 1 80 days after Allo -HSCT.Results 8 patients(28.6%)developed above grade 2 acute graft verse host disease(GVHD),2 patients developed grade 3 GVHD among the 8 patients.Two case with GVHD were cured by voriconazole with the therapeutic dose who occurred probably pulmonary invasive fungal infection at two months after Allo -HSCT.There were no other patients diagnosed fungal infection.No toxic efect were observed during the clinical observation during treatment with micafungin.5 patients appeared mild liver function abnormalities during treatment with voriconazole,and liver dysfunction were improved by symptomatic treatment.2 cases developed transient auditory hallucination and visual impairment induced by voriconazole.Conclusion Micafungin and reduced -dose voricon-azole are effective and safe prophylaxis in prevention early invasive fungal infection after HSCT.