Objective To research the expression,methylated regulation and clinical significances of miR-34b in chidren with acute leukemia(AL).Methods The methylation status of miR-34b promoter CpG islands were detected with methylation-specific polymerase chain reaction (MSP) in patients with AL.Then the expression of miR-34b was compared,which was detected by Taqman real-time fluorescence quantitative polymerase chain reaction (qRT-PCR),between the AL patients and normal group,in order to analyze their relationship with the clinical indicators.Resuits In 8 leukemia cell lines (U937,HL-60,MV4-11,M2R,K562,Raji,CCRF,DAMI) showed methylation,the positive rate of the methyl was 100％.Thirty-one acute lymphocytic leukemia(ALL) pediatric patients were newly diagnosed,and 24 cases showed methylation,the methylation positive rate 77.42％ (24/31).Nineteen acute myeloid leukemia(AML) patients were newly diagnosed,and 8 cases showed methylation,the methylation positive rate was 42.11％ (8/19 cases).There was no methylation in the 23 cases of normal children.The relative expression levels of miR-34b in the normal group,the group of leukemia cell lines,the group of ALL pediatric patients with newly diagnosed,the AML group and the group with mixed lineage leukemia gene rearrangement MLL+ were 5.22 ± 1.15,0.03 ± 0.03,1.65 ± 0.69,0.18 ± 0.06,0.64 ± 0.34,respectively.The findings indicated that there were significant differences in the relative expression levels of miR-34b between the normal group and the group of leukemia cell lines,the ALL group,the AML group,and the MLL+ group.The relative expression and methylated level of miR-34b had no statistically difference in gender,age at diagnosis,WBC count,chromosome,fusion gene,MLL gene rearrangement,and the minimal residual disease(LDH) levels in newly diagnosed AML patients(all P ＞ 0.05).And the relative expression and methylated level of miR-34b had no statistically difference in gender,age at diagnosis,WBC count,immunophenotype,chromosome,fusion gene,MLL gene rearrangement,TEL/AML1 gene,risk stratification,the minimal residual disease (MRD) in thirty-three days and the LDH levels in newly diagnosed ALL patients(all P ＞ 0.05).But as for the response to prednisone experiment,there was a significant difference between the sensitive group and the non sensitive group(P ＜ 0.05).Conclusions The expression level of miR-34b in AL was significantly lower and it was regulated by methylation mechanism,which implies that miR-34b may play a role of a tumor suppressor gene in the pathogenesis of leukemia.MiR-34b may affect the early treatment response of ALL patients,and it may be an indicator of risk stratification and poor prognosis in pediatric ALL.

Objective To investigate the predictive value of IL-9 cytokines in the patients with acute respiratory distress syndrome (ARDS).Methods According to Berlin definition of ARDS published in 2012,data of 28 patients with ARDS and another 22 healthy subjects as control were collected for prospective study from June,2013 to July,2014.Of them,there were 23 patients with severe pneumonia,1 patient with acute mercury poisoning,2 patients with severe acute pancreatitis,2 patients with acute paraquat poisoning.The survivors of ARDS patients were followed up.The ARDS patients were divided into moderate group (n =18) and severe group (n =10) as per the severity of the disease diagnosed at the first day after admission.And the ARDS patients were also divided into non-survival group (n =15) and survival group (n =13) according to the ARDS patients survived for 28 days.Three mLs of peripheral venous blood were collected in the early morning from fasted ARDS patients on the first and the third day after diagnosis of ARDS confirmed,and those of healthy subjects were collected on the first day after admission.The IL-9 cytokine level of peripheral venous blood detected by using enzyme linked immunosorbent assay (ELISA).The comparisons of levels of IL-9 cytokine were carried out between ARDS group and control group on the first day after diagnosis of ARDS established,between moderate group and severe group on the first day and the third day,and between survival group and non-survival group.The receiver operating characteristic (ROC) curve was used to evaluate the performance of IL-9 as a prognostic indicator in the early stage of ARDS.Data were analyzed by using SPSS 19.0 software.Results On the first day after diagnosis of ARDS,there were no statistically significant differences in age,APACHE Ⅱ score,procalcitonin (PCT),C-reactive protein (CRP),white blood cell count,lactate,and albumin between survival group and non-survival group (P ＞ 0.05).PH value in non-survival group was significantly lower than that in survival group (P＜0.05).IL-9 cytokine level of peripheral venous serum in ARDS group was significantly higher than that in healthy control group (P ＜ 0.05).There were no statistically significant differences in IL-9 level of peripheral venous serum both between moderate group and severe group and between survival group and non-survival group (P ＞ 0.05).On the third day,IL-9 level in severe group was significantly higher than that in moderate group (P ＜ 0.05),and that in survival group was significantly lower than that in non-survival group (P ＜ 0.05).The ROC of IL-9 at the first day for predicting mortality had all area under curve (AUC) to be 0.579 (95％ CI 0.361-0.798,P ＞ 0.05).The ROC of IL-9 on the third day for predicting mortality had AUC of 0.769 (95％ CI 0.592-0.947,P ＜ 0.05).When the cut-off value of IL-9 for the death followed up for 28 day' s was 2.88 pg/mL,the sensitivity was 86.7％,and the specificity was 61.5％.Conclusions IL-9 levels of in patients with ARDS were significantly higher,and IL-9 level can be helpful for the assessment of ARDS severity in the early stage,and for prognosis as well.

Objective To explore influential factors of local therapeutic effect in CT guided brachytherapy of 125I seeds for non-small-cell lung carcinoma (NSCLC).Methods Totally 141 primary NSCLC patients diagnosed by bronchoscope or puncture biopsy were treated with CT guided 125I seeds implantation treatment from 2003 January to 2005 January.Among them,26 patients were treated with seeds implantation only and remaining 115 combined with chemical therapy.Preplans were performed by using treatment planning system before the implantation.We took the implantation with the prescription dose of 80-110 Gy,1 seed per 1 cm3,under the guide of computed tomography.Six months after implantation treatment,CT graphs were taken to evaluate the therapeutic effect.Results All the patients were survival until 6 months after implantation,and 37 were complete remission,93 were partial remissions.The effective rate was 92.2％.Among all the observed factors,pathologic type (F =5.162,P =0.023),dose of cover 100％ tumor(D100) (F =100.713,P =0.000) and treatment methods (F =16.205,P =0.000) were the independent influent factors (P ＜ 0.05).Among these,D100 was the most important factor (P =0.000).Single factor analysis indicated that pathologic type (x2 =7.313,P =0.007),D100 (x2 =71.6,P =0.000)and treatment methods (x2 =20.5,P =0.000) were significant influent factors.Of all 141 cases,24 had complications during or after implantation treatment,while no severe complications were reported.There was no significant correlation between complication and local therapeutic effect (P ＞ 0.05).Conclusion CT guided implantation of 125I seeds for lung cancer has good clinical effects and few complications.D100 is the most important factor to influence the local therapeutic effect.Implantation treatment combined with chemotherapy is an ideal measure for NSCLC treatment.

OBJECTIVETo study the expression level and CpG island methylation status of miR-34b in leukemia cell lines and to research the effect of DNA methylation on the proliferation of leukemia cells regulated by miR-34b.

METHODTaqman real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was carried out to detect the relative expression of miR-34b in control group (bone marrow cells of 20 children without blood disease) and 8 leukemia cell lines (U937, HL-60, MV4-11, M2R, K562, Raji, CCRF, DAMI). Methylation-specific polymerase chain reaction (MSP) was carried out to detect the methylation differences of miR-34b in control group (Bone marrow cells of 23 children without blood disease), 8 leukemia cell lines. HL-60 and K562 were treated with methyltransferase inhibitor (5-aza-2-dC) for further detection of its methylation status and expression of miR-34b. Hsa-miR-34b mimics was transfected into K562 cell by liposome, the transfection efficiency was detected by flow cytometry. The cell proliferation of hsa-miR-34b transfected group in each stage was measured with CCK-8 assay, and then compared with non-transfected group and negative control group.

RESULTThe relative expression level of miR-34b in the group of children without blood disease and the group of leukemia cell lines were 5.22 ± 1.15, 0.03 ± 0.03. The results showed that, the group of leukemia cell lines was significantly different from the control group (t = 4.538, P < 0.01) . Eight leukemia cell lines showed methylation, the positive rate of the methyl was 100%. There was no methylation in the 23 cases of control group. After leukemia cell lines HL-60 and K562 were treated with 5-aza-2-dC, the methylated bands became obviously weakened, and the relative expression levels of miR-34b substantially increased 49.5 times and 18.8 times respectively. After hsa-miR-34b mimics was transfected into K562 cell by liposome, its transfection efficiency detected by flow cytometry was 61% and the cell proliferation was measured with CCK-8 assay from which it was found that the cell proliferation was significantly suppressed compared with the control group at 48 h (t = 9.303, P < 0.01), 72 h (t = 65.617, P < 0.01), 96 h (t = 36.878, P < 0.01) and 120 h (t = 18.748, P < 0.01) in hsa-miR-34b transfected group, with the inhibition rate of 12.2% (48 h), 45.7% (72 h), 32.5% (96 h) and 22.9% (120 h).

CONCLUSIONThe hypermethylation of promoter leads to decrease in the expression levels of miR-34b in leukemia cell lines, which attenuate mechanism of proliferative inhibition may be one of the reasons of occurrence or development of childhood leukemia.

Azacitidine ; Cell Line, Tumor ; Cell Proliferation ; genetics ; CpG Islands ; DNA Methylation ; HL-60 Cells ; Humans ; Leukemia ; genetics ; MicroRNAs ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction ; Transfection

OBJECTIVETo investigate the status of job burnout and its main influential factors in seafarers and to provide a scientific basis for ensuring the physical and psychological health of seafarers and increasing their working performance.

METHODSA total of 1027 seafarers, who underwent physical examination at Fujian International Travel Health Care Center from January to June, 2013, and left and entered China through the Fujian port, were selected. The status of job burnout was investigated using a job burnout scale. A total of 1027 questionnaires were sent out, and 989 valid ones (96.30%) were returned.

RESULTSThe scores of emotional exhaustion and cynicism were the highest in the youngest age group (<30 years), divorced or widowed group, or those with a monthly income per person over 10,000 yuan (P < 0.05). The score of reduced personal accomplishment was the highest in seafarers with a degree of junior high school or less or those with a monthly income per person of 3 000-6 000 yuan (P < 0.05). The highest scores of emotional exhaustion and cynicism were also seen in seafarers with the highest frequency of overtime working, high occupational stress, less than 6 hours' sleep per day, or poor sleep quality (P < 0.05). The highest score of reduced personal accomplishment was also seen in seafarers with the latest sail time lasting for more than six months, low occupational stress, or good sleep quality (P < 0.05). Multivariate analysis showed that poor sleep quality and occupational stress were the main risk factors for job burnout in seafarers, while physical exercise was a protective factor.

CONCLUSIONJob burnout among seafarers is influenced by many factors. Therefore, measures should be taken by relevant administrative departments and seafarers themselves to reduce the incidence of job burnout.

Adolescent ; Adult ; Burnout, Professional ; Female ; Humans ; Job Satisfaction ; Male ; Middle Aged ; Ships ; Surveys and Questionnaires ; Young Adult

Objective To evaluate the predictive role of TEL/AML1 fusion gene in protocol CCLG-ALL-2008 and to identify relevant factors influencing the outcome of ALL with TEL/AML1 fusion gene. Methods Ninety-nine patients with ALL harboring TEL/AML1 fusion gene (positive) and 329 cases without any specific fusion genes (negative) at diagnosis of B-lineage ALL from June 2008 to December 2014 were enrolled and their clinical and biological features were analyzed. Following-up ended in October 2015, the survival status was calculated by K-M curve and prognostic factors were analyzed by COX model. Results There were no differences between the two groups in age, white blood cell at the diagnostic stage, and treatment responses at 4 time points, namely, prednisone good response on day 8, M3 status of BM on D15, and the minimal residual disease (MRD) more than 1.0×10-3 on day 33 and 12th week. During the follow-up period, the relapse rate was lower in the positive group than that in the negative group (14/99 vs 69/329), the mortality rate of the negative group was twice of that in the positive group (55/329 vs 8/99). The five-year overall survival (OS) rate, relapse-free survival (RFS) rate and event-free survival (EFS) rate of the positive group were (86.1 ± 4.9)%, (80.7 ± 5.1)% and (78.9 ± 5.1)%, respectively, and (79 ±2.8)%, (72± 3.1)%, and (69.6+ 3.1)% for the negative group as well. COX regression analysis indicated that relapse and MRD level at the 12th week were independent prognostic factors on OS, RFS, and EFS (P<0.05) for the two groups. Conclusions TEL/AML1 fusion gene could be regarded as a relatively good indicator of risks in ALL children treated by CCLG-ALL-2008 protocol. ALL patients with TEL/AML1 are recommended to receive more intensive therapy including hematopoietic stem cell transplantation when the patients were high level of MRD on the 12th week after treatment.

Objective To investigate the expressions of programmed death-ligand 1 (PD-L1) and PD-L2 and phosphorylated protein kinase B (p-AKT) in diffuse large B-cell lymphoma (DLBCL) patients and their correlations with clinicopathological features and prognosis. Methods A total of 68 paraffin-embedded specimens of DLBCL patients diagnosed in Shanxi Provincial Cancer Hospital with detailed follow-up record from January 2010 to December 2012 were included in the study. The expressions of PD-L1, PD-L2 and p-AKT proteins in DLBCL were detected by using immunohistochemistry (IHC). Results The positive rate of PD-L1 protein in DLBCL patients was 22.1％ (15/68), which was related to germinal center B-cell (GCB) subtype or not (χ2= 5.591, P= 0.018), clinical stage (χ2= 3.969, P= 0.046), international prognostic index (IPI) grades (χ2=4.178, P=0.041) and treatment remission rate (χ2=6.587, P=0.010). The positive rate of PD-L2 protein in DLBCL patients was 14.7％ (10/68), which was related to extranodal metastasis or not (χ2=6.772, P= 0.009). The positive rate of p-AKT for DLBCL patients was 61.8％ (42/68), which was correlated with age (≥60 years old) or not (χ2=6.227, P=0.013), Eastern Cooperative Oncology Group (ECOG) grades (χ2=4.005, P=0.045), B symptoms (χ2=10.187, P=0.001) and treatment remission rate (χ2=4.096, P=0.043). Univariate survival analysis showed that the overall survival (OS) rate and progression free survival (PFS) rate of PD-L1 protein positive expression group were lower than those of PD-L1 protein negative expression group (both P< 0.05). In the patients with non-GCB subtype, OS rate and PFS rate of PD-L1 protein positive expression group were lower than those of PD-L1 protein negative expression group (both P<0.05). p-AKT protein positive expression group had poorer OS rate and PFS rate compared to p-AKT negative expression group (both P< 0.05). Correlation analysis showed that PD-L1 protein expression was correlated with PD-L2 and p-AKT proteins expressions (r= 0.380, P= 0.001;r= 0.273, P= 0.025). The prognosis was worse when p-AKT and PD-L1 proteins was co-expressed (P< 0.05). Multivariate analysis suggested high expressions of PD-L1 and p-AKT proteins were independent prognosis risk factors in DLBCL (both P<0.05). Conclusions The expressions of PD-L1 and p-AKT proteins may be involved in the occurrence and development of DLBCL. Blocking PD-1 and PD-L1 access or combined blocking could provide a promising future for the clinical therapy.

Objective:A rare case of δ-globin gene (HBD) mutation in Guangdong Province was analyzed to provide reference for avoiding misdiagnosis of δ-thalassemia in clinic.Methods:The patient was admitted to Guangdong Maternal and Child Health Hospital, and the peripheral blood sample was collected for hematological phenotypes [mean erythrocyte volume (MCV), mean erythrocyte hemoglobin content (MCH), hemoglobin (Hb)] and Hb typing analysis. The routine deletion and mutation of α-thalassemia and β-thalassemia genes were analyzed by PCR-flow fluorescence hybridization. At the same time, DNA sequencing was used to analyze the type of HBD mutation.Results:The results of hematological phenotypes analysis showed that MCV was 87.9 fl, MCH was 29.3 pg, and Hb content was 140 g/L. The results of Hb typing showed that the contents of Hb F, Hb A 2, Hb A 2 variant, and Hb A were 0.4%, 1.3%, 0.6%, and 97.7%, respectively. No abnormality was found in α-thalassemia and β-thalassemia genes by routine deletion and mutation detection. According to DNA sequencing analysis, the patient had HBD: c.349 C>G variant. Conclusion:The low Hb A 2 content (reference value is 2.5% - 3.5%) in this case is due to the mutation of HBD, HBD: c.349 C>G variant is rare in Chinese population.