To explore the role of the Chk2 protein expression and DNA double strand breaks (DSBs) repair in low dose hyper-radiosensitivity (HRS)/increased radioresistance (IRR) of non-small cell lung cancer, A549 cells were subjected to irradiation at the dosage ranging from 0.05-2 Gy. Clonogenic survival was measured by using fluorescence-activated cell sorting (FACS) plating technique. Percentage of cells in M-phase after low doses of X-irradiation was evaluated by phospho-histone H3-FITC/PI and Western blotting was used to detect protein expression of Chk2 and phospo-Chk2. DNA DSBs repair efficiency was also measured by induction and persistence of γ-H2AX. The results showed that the killing ability of irradiation with A549 cells increased at low conditioning dose below 0.3 Gy. Within the dose of 0.3 to 0.5 Gy, A549 cells showed a certain extent of radiation resistance. And when the dose was more than 0.5 Gy, survival fraction exhibited a negative correlation with the dosage. There was no difference between the 0.1 or 0.2 Gy dosage groups and the un-irradiated group in terms of the percentage of cells in M phase. But in the high dosage group (0.3-1.0 Gy), the percentage of cells in M phase was decreased markedly. In addition, the percentage of cells in M phase began to decrease two hours after irradiation. One hour after irradiation, there was no conspicuous activation of Chk2 kinase in 0.1 or 0.2 Gy group, but when the irradiation dose reached 0.3 Gy or higher, Chk2 kinase started to be activated and the activation level showed no significant difference among high dosage groups (0.4, 0.5, 1.0 Gy). Within 1 to 6 h, the DNA DSBs repair efficiency was decreased at 0.2 Gy but increased at 0.5 Gy and 1.0 Gy, which was in line with Chk2 activation. We are led to conclude that the mechanism of HRS/IRR in A549 cell line was probably due to early G(2)/M checkpoint arrest and enhanced DNA DSBs repair. In this regard, Chk2 activation plays a key role in G(2)/M checkpoint activation.

To explore the role of the Chk2 protein expression and DNA double strand breaks (DSBs) repair in low dose hyper-radiosensitivity (HRS)/increased radioresistance (IRR) of non-small cell lung cancer, A549 cells were subjected to irradiation at the dosage ranging from 0.05-2 Gy. Clonogenic survival was measured by using fluorescence-activated cell sorting (FACS) plating technique. Percentage of cells in M-phase after low doses of X-irradiation was evaluated by phospho-histone H3-FITC/PI and Western blotting was used to detect protein expression of Chk2 and phospo-Chk2. DNA DSBs repair efficiency was also measured by induction and persistence of γ-H2AX. The results showed that the killing ability of irradiation with A549 cells increased at low conditioning dose below 0.3 Gy. Within the dose of 0.3 to 0.5 Gy, A549 cells showed a certain extent of radiation resistance. And when the dose was more than 0.5 Gy, survival fraction exhibited a negative correlation with the dosage. There was no difference between the 0.1 or 0.2 Gy dosage groups and the un-irradiated group in terms of the percentage of cells in M phase. But in the high dosage group (0.3-1.0 Gy), the percentage of cells in M phase was decreased markedly. In addition, the percentage of cells in M phase began to decrease two hours after irradiation. One hour after irradiation, there was no conspicuous activation of Chk2 kinase in 0.1 or 0.2 Gy group, but when the irradiation dose reached 0.3 Gy or higher, Chk2 kinase started to be activated and the activation level showed no significant difference among high dosage groups (0.4, 0.5, 1.0 Gy). Within 1 to 6 h, the DNA DSBs repair efficiency was decreased at 0.2 Gy but increased at 0.5 Gy and 1.0 Gy, which was in line with Chk2 activation. We are led to conclude that the mechanism of HRS/IRR in A549 cell line was probably due to early G(2)/M checkpoint arrest and enhanced DNA DSBs repair. In this regard, Chk2 activation plays a key role in G(2)/M checkpoint activation.
Adenocarcinoma ; genetics ; metabolism ; Cell Line, Tumor ; Checkpoint Kinase 2 ; genetics ; metabolism ; DNA Damage ; genetics ; DNA Repair ; genetics ; Humans ; Lung Neoplasms ; genetics ; metabolism ; Radiation Tolerance ; genetics

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 investigate the perinatal outcome , risk factors and long-term outcome of pregnancy complicated with pulmonary arterial hypertension (PAH) and congenital heart diseases (CHD). Methods Clinical data of 110 pregnant women who were diagnosed as PAH-CHD were retrospectively analyzed in the Department of Obstetrics and Gynecology and Surgical Intensive Care Unit at Beijing Anzhen Hospital from 2004 to 2013.The survival and treatment status were followed up .Results 110 subjects consisted of 11 mild PAH, 33 moderate and 66 severe ones .The incidences of deterioration in New York Heart Association ( NYHA ) classes (≥2 ) during pregnancy , respiratory failure , pulmonary hypertension crisis and arrhythmia were 25.5% (28/110),7.3% (8/110),10.0% (11/110),10.0% (11/110) respectively.Among them, the difference of deterioration in NYHA classes (≥2) during pregnancy among the three groups was statistically significant .A total of 8 ( 7.3%) maternal deaths occurred during hospitalization , all of whom were severe PAH cases .Multivariate analysis showed that pulmonary artery systolic pressure was a risk factor of perioperative death (OR=1.042, P=0.005).There were 55 cases (50.0%) of term delivery, and 35 cases (31.8%) of iatrogenic abortion.The proportion of term delivery in the severe PAH group was significantly lower . The proportion of iatrogenic abortion and small for gestational age infant ( SGA ) were higher in severe group .The incidence of neonatal malformations was 8.0%(6/75).The follow-up rate was 61.8%(63/102).Sudden death was reported in a parturient a few days after discharge .The remaining 62 patients survived during follow-up, while 53 patients (85.5%) were functional class ( FC ) Ⅰ -Ⅱ, 9 ( 14.5%) were FC Ⅲ -Ⅳ at follow-up.The cardiac function deterioration during pregnancy was not significantly correlated with long-term deterioration (P =0.767). Conclusions Perinatal mortality and the incidence of maternal and fetal adverse events were high in pregnancy with PAH-CHD.Pulmonary artery systolic pressure is a major risk factor for perioperative mortality in pregnant women .PAH-CHD woman had good overall outcome after puerperium .

OBJECTIVETo investigate the expression level of SET gene in patients with acute myeloid leukemia (AML) and evaluate its significance.

METHODSThe expression level of SET gene in 141 de novo AML patients was determined by real time quantitative PCR (RQ-PCR), and its relationship with the clinical features and outcomes of these patients were analyzed.

RESULTSSET gene transcript level was detected in 141 AML patients with the median expression level of 0.86(range 0.02-15.69). AML patients with higher SET gene expression had a higher level of white blood cell (WBC ≥ 100 × 10⁹/L) count than of lower SET gene expression ones (31.0% vs 11.4%, P=0.005). In the 136 patients who received treatment after diagnosis, higher SET gene expression group had lower complete remission rate (50.0%) than of lower expression cohort (73.5%) after two cycles of chemotherapy (P=0.005). Survival analysis showed that patients with higher SET gene expression had significantly shorter overall survival(OS) (10 months vs 22 months, P=0.001) and event-free survival (EFS) (2 months vs 14 months, P=0.005) than of lower SET gene expression ones. Multivariate COX regression analysis showed SET overexpression was an independent prognostic factor for OS. In the patients with the normal karyotype, higher SET expression group also had significantly shorter OS (12 months vs 35 months, P=0.010) and EFS (4 months vs 14 months, P=0.026) than of lower SET expression ones.

CONCLUSIONHigh expression of SET gene was associated with poor prognosis and might be a prognostic molecular marker of AML.

Disease-Free Survival ; Gene Expression Regulation, Neoplastic ; Histone Chaperones ; genetics ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Prognosis ; Remission Induction ; Transcription Factors ; genetics

Objective To establish a nucleic acid assay for detection of Paragonimus skrjabini based on the recombinase-aided isothermal amplification (RAA) technique, and to preliminarily evaluate its detection efficiency. Methods The metacercariae of P. skrjabini, P. westermani and Euparagonimus cenocopiosus were isolated from crabs, and genomic DNA was extracted for molecular characterization. The cytochrome coxidase 1 (cox1) gene sequence of P. skrjabini was selected as the target gene fragment, and the primers and probes were designed, screened and synthesized for RAA assay. The genomic DNA of P. skrjabini metacercariae from Jiyuan City and Yiyang County of Luoyang City, Henan Province were used as templates for verification of the fluorescent RAA assay. The fluorescent RAA assay was performed to detect different concentrations of plasmids containing target gene fragment and P. skrjabini metacercariae genomic DNA to determine the sensitivity. Fluorescent RAA assay was performed with recombinant plasmids containing P. skrjabini cox1 gene sequences at different concentrations and P. skrjabini genomic DNA as templates to evaluate its sensitivity, and the genomic DNA of P. westermani, E. cenocopiosus, Clonorchis sinensis and Schistosoma japonicum was detected with fluorescent RAA assay to evaluate its specificity. Results P. skrjabini, P. westermani and E. cenocopiosus metacercariae were isolated from crabs, respectively. Molecular characterization and phylogenetic analysis confirmed their homology with the genes sequences of standard Paragonimus strains in GenBank. A fluorescent RAA assay was successfully established for nucleic acid detection of P. skrjabini, and the genomic DNA of P. skrjabini metacercariae from Jiyuan City and Yiyang County of Luoyang City, Henan Province was amplified using the fluorescent RAA assay within 5 min, while the negative control was not amplified. If the recombinant plasmid containing P. skrjabini cox1 gene sequences was used as templates, the fluorescent RAA assay showed the lowest detection limit of 10 copies/μL, and positive amplification was observed within 5 min. If genomic DNA was used as templates, the fluorescent RAA assay showed the lowest detection limit of 10 pg/μL, and all positive amplifications were found within 5 to 10 min. In addition, the fluorescent RAA assay was tested negative for P. westermani, E. cenocopiosus, C. sinensis and S. japonicum. Conclusions A rapid, sensitive and specific fluorescent RAA assay is successfully established for nucleic acid detection of P. skrjabini, which has potential values in rapid field detection and species identification in freshwater crabs in areas endemic for P. skrjabini.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Humans ; Induction Chemotherapy ; Leukemia, Myeloid, Acute ; drug therapy ; Middle Aged

OBJECTIVETo explore the incidence of chromosome 1 abnormality in myelodysplastic syndrome（MDS）to couple its association with clinical presentation and prognosis.

METHODSR- band karyotype analyses were performed in 672 cases of MDS between 2010 and 2013. Clinical data of those with abnormal chromosome l were collected and then analyzed factors affecting the prognosis.

RESULTSOf 672 cases of patients with MDS, chromosome 1 aberration［der（1）, dup（1）, －1 were most frequent］ were found in 41（6.1%）cases. 1q trisomy was found in 18/41（43.9%）cases, and the most common patterns were duplication of the long arm as well as unbalanced translocation with other chromosomes. Of 41 patients with chromosomal 1 abnormality, 32 cases were accompanied with other chromosomal aberration, usually involving 3 or more abnormal chromosomal karyotypes, e.g., chromosome 8, 7 abnormalities. According to IPSS-R scoring system, 19 patients were diagnosed with very high risk, 10 patients high risk, 10 patients intermediate risk and 2 patients low risk MDS. 9 patients transformed into acute leukemia with median transforming time of 7.18（0.56-54.28）months. Median survival of 36 cases after 2010 was 17.48（95% CI 14.38-20.58）months. There were significant differences on median survival between RAEB and non-RAEB groups（χ²=10.398, P=0.001）, and between with more than 3 chromosome abnormalities and with less than 3 groups（χ²=3.939, P=0.047）. RAEB was identified as an independent risk factor for the prognosis of MDS with chromosome 1 abnormality.

CONCLUSIONChromosome 1 aberration was not rare in MDS. 1q trisomy was the most common abnormal karyotype in China, which often accompanied with other chromosomal abnormalities. The prognosis of MDS patients with chromosome 1 abnormality was poor, especially worse in those diagnosed with RAEB-1, RAEB-2 and with more than 3 chromosome abnormality. For patients whose percentage of bone marrow blasts less than 5%, the prognosis of patients with 1q trisomy was better than those without 1q trisomy. RAEB was identified as an independent risk factor for the prognosis of MDS with chromosome 1 abnormality.

Abnormal Karyotype ; Acute Disease ; Anemia, Refractory, with Excess of Blasts ; Bone Marrow ; China ; Chromosome Aberrations ; Chromosome Banding ; Chromosomes, Human, Pair 1 ; genetics ; Humans ; Karyotyping ; Leukemia ; diagnosis ; genetics ; Myelodysplastic Syndromes ; diagnosis ; genetics ; Prognosis ; Risk Factors ; Trisomy