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Objective To examine the influencing factors related to clinical efficacy and outcomes of adult primary immune thrombocytopenia (ITP).Methods The clinical data of 161 cases of ITP admitted in the Second Hospital of Shanxi Medical University from June 2013 to March 2017 were collected.The influencing factors related to clinical efficacy and prognosis of adult ITP patients were analyzed.Results There were 60 males and 101 females with a M/F ratio of 0.59∶1 and a median age of 45 years (18-84 years).There were 109 newly diagnosed ITP cases,14 persistent ITP cases and 38 chronic ITP cases in this series.Seventy nine patients received intravenous immunoglobulin g (IVIg) treatment and 82 patients received high dose-dexamethasone treatment.There were no significant differences in clinical efficacy [91.13%(72/79) vs.87.80%(72/82),x2=0.181,P=0.914] and relapse rate [36.11%(26/72) vs.30.55%(22/72),x2=0.189,P=0.910] between IVIg and high dose-dexamethosone groups.Multivariate regression analysis showed that bleeding score ≥2 was the independent risk factor for the lower clinical efficacy (RR=1.415,95%CI:1.008-1.986,P<0.05).Patients were followed up for a median of 9.0 months (0.5-55.0 months),48 patients relapsed with a relapse rate of 33.33% and a median relapse time of 1.8 months (0.5-24.0 months).Conclusions IVIg and high dose-dexamethasone have the similar clinical efficacy and relapse rate for treatment of adult ITP.The patients with the bleeding score ≥2 are more likely to get lower remission rate.
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Objective To analyze the clinical features and risk factors for an emerging infection during the first induction chemotherapy in elderly patients with acute leukemia.Methods A retrospective analysis of clinical data of 79 elderly patients with newly diagnosed acute leukemia was performed in Second Hospital of Shanxi Medical University from January 2014 to May 2016.Results The 70 cases among 79 elderly patients with acute leukemia were suffered from infection with infection incidence rate of 88.6% (70/79)during first induction chemotherapy.The infection-related fatality rate was 8.6 % (6/70).Being clear about sites of infection accounted for 90.0 % (63/70),and the top three infection sites were the lungs,gastrointestinal tract and the bloodstream.113 pathogenic strains were detected,including gram-negative bacilli accounting for 42.5 % (48/113),Gram-positive cocci for 30.1% (34/113),fungi for 24.8% (28/113),the virus for 2.7% (3/113).Based on clinically and confirmatively diagnosis,the invasive fungal diseases mostly as Candida accounted for 30.4 % (24/79),mixed infections accounted for 34.3% (24/70).Univariate analysis showed agranulocytosis and AML were risk factors for infection.Logistic multivariate regression analysis showed that agranulocytosis was a risk factor for infection (OR=12.010,95%CI:2.346-107.973,P=0.000).The infection does not affect a complete remission rate of acute leukemia (x2 =0.001,P=0.983).Conclusions For newly diagnosed elderly acute leukemia patients,an emerging infection during the first induction chemotherapy is characterized by a high incidence,high fungal infection rate,most common site in lung,Gram-negative bacteria as most common pathogen,and an increased infection rate by agranulocytosis.The infection does not affect the remission rate of acute leukemia.
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Objective@#To investigate the effect of biology and mTOR pathway activity of down-regulated TSC2 gene expression on U937 leukemia cells.@*Methods@#Gene expression was down-regulated by lentivirus induced RNA interference on TSC2 high expressed U937 cell line; the proliferation, apoptosis and differentiation were detected by CCK-8 assay, colony formation assay and flow cytometry; the gene expression level and protein kinase activity were detected by qRT-PCR and Western blot.@*Results@#Down-regulated expression of TSC2 gene promoted U937 cell proliferation and colony formation ability (P<0.05) . The proportion in G0/G1 phase of TSC2 down-regulated U937 cell was much lower than that of the control cells [ (52.53±3.75) % vs (75.10±4.33) %, t=6.829, P=0.002], the S phase [ (22.43±1.00) % vs (15.47±1.20) %, t=-5.581, P=0.019] and G2/M phase [ (25.03±4.34) % vs (14.33±0.91) %, t=-5.413, P=0.013] was remarkably higher than that of the control cells (P<0.05) . There were no statistically significant differences in cell apoptosis and differentiation (P>0.05) . Down-regulation of TSC2 led to the increased activity of mTOR, 4EBP1 and S6K1, but did not influence the activity of AKT. The expressions of proliferation related cyclinD1, c-myc and PTEN were also up-regulated after TSC2 silenced, but the expressions of P27KIP and BCL-XL were not changed.@*Conclusion@#Downregulation of TSC2 could promote the proliferation of U937 cells through up-regulation of mTOR activity.
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Objective To investigate the mutations of epigenetic regulation factor ASXL1 gene in myelodysplastic syndrome(MDS).Methods Mutation analysis of ASXL1 gene in 53 de novo MDS patients and 20 healthy persons was performed by using polymerase chain reaction(PCR)followed by sequence analysis at DNA level.The clinical and laboratory characteristics were compared in MDS patients with ASXL1 gene mutation and ASXL1 wild type.ASXL1 mutation in mRNA level was detected by using reverse transcription PCR(RT-PCR)followed by sequence analysis.Results ASXL1 gene mutations were observed in 9 cases(16.9%)of 53 MDS patients.6 mutation types were detected,including 4 frameshift mutations types(2 cases with p.Glu635ArgfsX15,3 cases with p.Gly646TrpfsX12,1 case with p.Ala640GlyfsX14 and 1 case with p.Gly790TrpfsX10)and 2 nonsense mutation types(1 case with p.Gln1063X and 1 case with p.Gln695X).All the mutations were heterozygous,and p.Gly790TrpfsX10 and p.Gln695X were new mutation types.In addition,a single nucletide polymorphism(SNP)p.Gly652Ser was also detected in 4 cases with MDS.5 cases of p.G652S SNP and 1 case of p.Leu1173Leu SNP were detected in 20 healthy people.Frameshift mutation(p.Gly646TrpfsX12)could be detected at mRNA level by using RT-PCR.Differences were not observed in red blood cell counts,white blood cell counts,platelet counts,hemoglobin levels,reticulocyte,neutrophil granulocyte,the peripheral blood lymphocytes percentage,T-cell subsets in the peripheral blood,the proportion of primitive cell in the marrow and MDS types between the patients with ASXL1 gene mutation and ASXL1 wild type patients(P >0.05).Conclusion There is a high frequency of ASXL1 gene mutation in MDS patients,which can be detected at mRNA level.
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Objective To investigate the mutations of epigenetic regulation factor ASXL1 gene in myelodysplastic syndrome(MDS).Methods Mutation analysis of ASXL1 gene in 53 de novo MDS patients and 20 healthy persons was performed by using polymerase chain reaction(PCR)followed by sequence analysis at DNA level.The clinical and laboratory characteristics were compared in MDS patients with ASXL1 gene mutation and ASXL1 wild type.ASXL1 mutation in mRNA level was detected by using reverse transcription PCR(RT-PCR)followed by sequence analysis.Results ASXL1 gene mutations were observed in 9 cases(16.9%)of 53 MDS patients.6 mutation types were detected,including 4 frameshift mutations types(2 cases with p.Glu635ArgfsX15,3 cases with p.Gly646TrpfsX12,1 case with p.Ala640GlyfsX14 and 1 case with p.Gly790TrpfsX10)and 2 nonsense mutation types(1 case with p.Gln1063X and 1 case with p.Gln695X).All the mutations were heterozygous,and p.Gly790TrpfsX10 and p.Gln695X were new mutation types.In addition,a single nucletide polymorphism(SNP)p.Gly652Ser was also detected in 4 cases with MDS.5 cases of p.G652S SNP and 1 case of p.Leu1173Leu SNP were detected in 20 healthy people.Frameshift mutation(p.Gly646TrpfsX12)could be detected at mRNA level by using RT-PCR.Differences were not observed in red blood cell counts,white blood cell counts,platelet counts,hemoglobin levels,reticulocyte,neutrophil granulocyte,the peripheral blood lymphocytes percentage,T-cell subsets in the peripheral blood,the proportion of primitive cell in the marrow and MDS types between the patients with ASXL1 gene mutation and ASXL1 wild type patients(P >0.05).Conclusion There is a high frequency of ASXL1 gene mutation in MDS patients,which can be detected at mRNA level.
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Objective To explore the relationship between T-cell acute lymphoblastic leukemia and the Notch signaling pathway.Methods Human T-cell acute lymphoblastic leukemia SupT1 cells were infected with the lentiviral vector made up specific Notch1-shRNA gene and nonspecific Notch1-shRNA gene.The inhibitive rate of SupT1 cells was detected by CCK-8.The rates of early apoptotic cells (Annexin V+/ 7-AAD-) and late apoptotic cells (Annexin V+/7-AAD+) were analyzed by flow cytometry and the expression levels of Notch1 receptor gene and downstream target genes were assessed by quantitative reverse transcription and polymerase chain reaction (QT-PCR).Results The cell inhibition rates of Notch1 interference group,control group and empty vector group at 96 h were 0.902±0.013,0,and 0.486±0.084,respectively,and it was increased obviously in Notch1 interference group (both P < 0.05).The cell early apoptosis rates of the three groups were (15.27±0.31) %,(5.57±0.25) %,(5.80±0.20) %,respectively.The cell early apoptosis rate of Notch1 interference group was increased obviously compared with the control group and empty vector group (both P < 0.05).While the cell late apoptosis rates had no significant difference among the three groups (P > 0.05).The mRNA expression levels of Notch1 receptor gene and its target genes (Hesl,c-myc,NF-κB) at 48 h,72 h and 96 h were higher than those in the control group and empty vector group (all P < 0.05).Conclusions The specificity of Notch1-shRNA can effectively decrease the Notch1 mRNA expression,and reduce the expression level of downstream target genes.Notch1 cut can inhibit the proliferation of SupT1 cells,and promote the early apoptosis.
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Objective To investigate the relevance between CD25 and common markers of immune phenotype in acute myeloid leukemia (AML), and the relationship between CD25 and FLT3-ITD gene. Methods The expression of mono-clonal antibodies, including, CD7, CD117, CD33, CD34, CD38, cyMPO, CD13, CD36, CD56, CD11b, CD19, CD16, CD14, CD64 and interleukin-2 three receptor chain CD25 (αchain), CD122 (β chain), CD132 (γ chain) were detected by immunofluorescence flow cytometry in 36 cases of AML bone marrow blast. The expression of every group of pa-tients with FLT3-ITD gene were detected by PCR. Results In 36 cases of AML patients, the rate of CD25+was 13.89%(5/36), 12.5% (1/8) in AML-M2 group, 18.18% (2/11) in AML-M4 group, 20% (2/10) in AML-M5 group, respectively, and expression rate of CD25+had been no statistically differences between those subgroups (P> 0.05). The percentage of bone marrow blasts which express CD7, CD117, CD33, CD34, CD38, cyMPO, CD13, CD36, CD56, CD11b, CD19, CD16, CD14, CD64, CD122, CD132 monoclonal antibodies has no significant difference between CD25+AML-M2 group and CD25-AML-M2 group, CD25+AML-M4 group and CD25-AML-M4 group, CD25+AML-M5 group and CD25-AML-M5 group, respectively (P> 0.05). Seven cases of AML patients with FLT3-ITD mutation, in five cases of CD25+AML pa-tients three case accompany FLT3-ITD+ mutation, the rates of FLT3-ITD+in CD25+AML was 60%, and higher than CD25-AML group (P <0.05). Conclusion (1)The expression rate of CD25 in AML patients is lower, and no significant correlation in patients in different AML FAB subtypes and common myeloid phenotype markers. (2)Since the FLT3-ITD mutation rates are higher in CD25+AML patients, there is important significance to clear the correlation between CD25 and FLT3-ITD gene.
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Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity. CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Nobiletin potently activated CFTR chloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTR(inh)-172 could completely reverse the effect. Preliminary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may be a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis.
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Recently, more research about the plant bioreactor expressing genes encoding human proteins was reported. In the present study, the cDNA of the human gene keratinocyte growth factor 2 (KGF2) was replaced with plant preferred codons by PCR, and the modified full-length cDNA was cloned into the plant expression vector pCAMBIA-YO containing the oil-body promoter. The fusion construct pCAMBIA-YO-KGF2 was transformed into Brassica napus by Agrobacterium tumefacien-mediated cotyledon transformation method. The transgenic seedlings were identified by PCR, Southern and western blot analysis all showed that KGF2 gene was successfully expressed in in transgenic Brassica napus.