Relationship between DNMT1 and Methylation of SHP-1 Promoter 2 in K562 Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of DNA methyltransferase 1 ( DNMT1 ) with hematopoietic cell phosphatase (SHP-1) gene expression and promoter 2 methylation status in cell line K562.</p><p><b>METHODS</b>The promoter sequence of SHP-1 gene promoter 2 in NCBI database was analyzed, the K562 cells were transfected with the lentiviral plasmids-the specified retroviral vector psiHIV-mU6-shDNMT1 and psiHIV-mU6-mcherryFP-control. The methylation status of SHP-1 gene promoter 2 in K562 cells was detected by methylation-specific polymerase chain reaction (MSP) and bisulfite-modified sequencing (BSP). Western blot was used to detect the protein expression level of SHP-1 and DNMT1, the SYBR Green fluorescence quantitative PCR was used to detect the expression of SHP-1 mRNA.</p><p><b>RESULTS</b>It was found that the promoter 2 of SHP-1 gene located between -577 bp to +300 bp, and 22 CpG sites contained between -353 bp-+182 bp were aberrantly hypermethylated and the SHP-1 could not be detected in K562 cells. In vitro, the detection demonstrated that the expression level of DNMT1 in K562 cells transfected with psiHIV-mU6-shDNMT1 was 0.48±0.06 significantly lower than that of psiHIV-mU6-control group (1.33±0.19)(t= 4.18, P<0.05). The expression of SHP-1 mRNA in K562 cells transfected with psiHIV-mU6-shDNMT1 was significantly higher than that in K562 cells transfected with psiHIV-mU6-shDNMT1 (14.23±3.83 vs 1.031±0.156)(P<0.01). DNMT1 silencing induced demethylation of the 22 CpG sites located in the SHP-1 promoter 2, and SHP-1 gene was re-expression in K562 cells.</p><p><b>CONCLUSION</b>The DNMT1 in K562 cells relates with the hypermethylation and silencing of SHP-1 promoter in K562 cells.</p>

Current Status of Community-Acquired Pneumonia in Patients with Chronic Obstructive Pulmonary Disease / 中华医学杂志(英文版)

<p><b>Objective</b>Worldwide, community-acquired pneumonia (CAP) is a common infection that occurs in older adults, who may have pulmonary comorbidities, including chronic obstructive pulmonary disease (COPD). Although there have been clinical studies on the coexistence of CAP with COPD, there remain some controversial findings. This review presents the current status of COPD in CAP patients, including the disease burden, clinical characteristics, risk factors, microbial etiology, and antibiotic treatment.</p><p><b>Data Sources</b>A literature review included full peer-reviewed publications up to January 2018 derived from the PubMed database, using the keywords "community-acquired pneumonia" and "chronic obstructive pulmonary disease".</p><p><b>Study Selection</b>Papers in English were reviewed, with no restriction on study design.</p><p><b>Results</b>COPD patients who are treated with inhaled corticosteroids are at an increased risk of CAP and have a worse prognosis, but data regarding the increased mortality remains unclear. Although Streptococcus pneumoniae is still regarded as the most common bacteria isolated from patients with CAP and COPD, Pseudomonas aeruginosa is also important, and physicians should pay close attention to the occurrence of antimicrobial resistance, particularly in these two organisms.</p><p><b>Conclusions</b>COPD is a common and important predisposing comorbidity in patients who develop CAP. COPD often aggravates the clinical symptoms of patients with CAP, complicating treatment, but generally does not appear to affect prognosis.</p>

Expression of SHP-1 mRNA in Patients with Myelogenous Leukemia and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of SHP-1 mRNA in patients with myelogenous leukemia.</p><p><b>METHODS</b>The SYBR Green-based qRT-PCR was used to assess SHP-1 mRNA levels in 54 patients with chronic myelogenous leukemia (CML), 30 cases of de novo acute myelogenous leukemia (AML) and 10 persons without malignancy as controls.</p><p><b>RESULTS</b>The relative expression levels of SHP-1 mRNA in control group (CG), chronic phase CML (CP-CML) group, advanced phase of CML (including accelerated phase CML and blastic phase CML) group and AML group were 1.15±0.62, 4.96±1.76, 2.60±0.90 and 0.45±0.20, respectively. The expression of SHP-1 mRNA in patients with CML significantly increased in comparison with that in CG(P<0.05). Meanwhile, the expression of SHP-1 mRNA in CP-CML group very significantly increased as compared with that in advanced stage of CML group(P<0.0001). The expression of SHP-1 mRNA in AML group significantly decreased as compared with that in CG group(P=0.0442). In CP-CML group, statistical analysis showed that SHP-1 mRNA expression at baseline in optimal responders (5.712±0.4476) was significantly higher than that in the suboptimal or failed responders (4.044±0.3701)(P=0.0090). Meanwhile, the SHP-1 mRNA expression in AML patients was higher than that in CR group (0.4984±0.05164) and non-CR group (0.3537±0.02388)(P=0.0017).</p><p><b>CONCLUSION</b>The SHP-1 mRNA levels in CML patients are higher than that in AML patients, and probably correlats with disease progression of CML. The mRNA expression level of SHP-1 may be a molecular marker to predict early response to inatinib treatment in CP-CML and AML.</p>

Overexpression of SHP-1 Enhances the Sensitivity of K562 Cells to Imatinib / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of overexpression of SH2-containing tyrosine phosphatase 1 (SHP-1) on sensitivity of chronic myelogenous 1eukemia (CML) K562 cell line to imatinib and its related mechamism.</p><p><b>METHODS</b>K562 cells were infected with the lentiviral plasmids containing the specified retroviral vector (pEX-SHP-1-puro-Lv105) or the mock vector (pEX-EGFP-puro-Lv105). The expression of SHP-1 in K562 cells treated with 0.2 µmol/L imatinib (IM) for 72 h was determined by Western blot. After transfection the CCK-8 assay was used to determine the proliferation of the tramfected K562 cells (K562(SHP-1) and K562(EGFP) cells) at 72 h after exposure to different doses of IM, the half inhibitary concentration (IC50) was calculated. The mechanisms of the overexpression effects of SHP-1 and IM on the proliferation in K562 cells was investigated, the BCR-ABL1 activity and the level of tyrosine phosphorylation of CrkL (pCrkL) was measured by flow cytometry; the Western blot was used to detect the expression and activity of these molecules controlling cell growth, including MAPK, AKT, STAT5 and JAK2.</p><p><b>RESULTS</b>After exposure of K562 cells to 0.08 µmol/L IM for 72 h, there was no significant change of SHP-1 expression in K562 cells. After exposure to 0.2 µmol/L of IM for 72 h, the inhibitory rate of K562(SHP-1) group was higher than that of K562(EGFP) group (P < 0.05), indicating that overexpression of SHP-1 in K562 cells could enhance the proliferation inhtibition effect of IM on K562 cells. The IC50 of IM in K562(SHP-1) cells was the lower as compared with that of K562(EGFP) cells (P < 0.05) after exposure to different concentrations of IM for 72 h. The slope of K562(SHP-1) cells was the largest ranging 0.02 - 0.16 µmol/L of IM. Overexpression of SHP-1 and IM could inhibit the activity BCR-ABL1, MAPK, AKT, STAT5 and JAK2 signaling pathways in the K562 cell line and displayed a synergistic effect.</p><p><b>CONCLUSION</b>SHP-1 inhibits BCR-ABL1, MAPK, AKT, STAT5 and JAK2 signaling pathways in K562 cells, the overexpression of SHP-1 can enhance the sensitivity of K562 cells to IM.</p>

Expression and Clinical Significance of DNMT in Patients with Chronic Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of DNA methyltransferases (DNMT) mRNA in patients with chronic myeloid leukemia (CML).</p><p><b>METHODS</b>The expression levels of DNMT mRNA in mononucllear cells (MNC) of bone marrow or in peripheral blood of 93 CML patients in 3 different phases and 10 normal controls (NC) were detected by SYBR Green flurescent quatitative PCR.</p><p><b>RESULTS</b>The relative expression levels of DNMT1 mRNA in NC, chronic phase CML (CML-CP), accelerated phase (CML-AP) and blastic phase (CML-BP) were 1.45 ± 0.22, 1.83 ± 0.63, 2.95 ± 0.87 and 3.24 ± 1.39 resectively. The expression of DNMT1 mRNA showed no statistically significant difference between CML-CP and NC (P = 0.28). The expression of DNMT1 mRNA in advanced stages (including CML-AP and CML-BP) of CML obviously increased in comparison with CML-CP and NC (P < 0.05). The expression of DNMT1 mRNA in CML-AP was not significantly different from that in CML-BP (P = 0.336). The relative expression levels of DNMT3a mRNA in NC, CML-CP, CML-AP and CML-BP groups were 1.29 ± 0.34, 1.34 ± 0.46, 2.33 ± 1.05 and 3.18 ± 1.23 resectively. And the expression levels of DNMT3a mRNA were not statistically significantly different between CML-CP and NC (P = 0.844). The results showed that the expression of DNMT3a mRNA in the advanced phase of CML significantly increased in comparison with that in CML-CP and NC (P < 0.05). Meanwhile, the expression of DNMT3a mRNA in CML-AP was not different from that in CML-BP (P = 0.304). The relative expression levels of DNMT3b mRNA in NC, CML-CP, CML-AP and CML-BP groups were 1.37 ± 0.31, 16.41 ± 22.50, 9.36 ± 5.50 and 12.17 ± 13.44 resectively. It was also found that the level of DNMT3b mRNA in CML significantly increased in comparison with NC (P < 0.05), and that the between the 3 different phase of CML was not statistically significantly different (P >0.05).</p><p><b>CONCLUSION</b>The expression of DNMT mRNA increases in advanced CML as compared with normal controls and CML-CP, and the increased levels of DNMT mRNA probably correlate with disease progression in CML.</p>

Effects of CRF receptor antagonist on rem sleep in neonatal rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the role of NB127914, a CRF R1 receptor antagonist, in the regulation of neonatal sleep/wake cycle.</p><p><b>METHODS</b>Rat pups were surgically implanted with electrodes at postnatal day(PN) 13. At PN 14, 6 hours polysomnographic recording data were continuously collected before and after administration of various doses of NBI 27914, atropine and the same amount of saline.</p><p><b>RESULTS</b>Compared with baseline, rapid eye movement (REM) sleep was significantly reduced and was replaced primarily by non-REM (NREM) sleep in all groups treated with NBI, but not with dimethyl sulfoxide/saline. Atropine suppressed REM sleep significantly and increased wakefulness simultaneously.</p><p><b>CONCLUSION</b>Blockage of corticotropin-releasing factor (CRF) R1 receptors deprives neonatal rat REM sleep.</p>

Evaluation on the indirect economic burden of stroke using combination of disability-adjusted life years and human capital method / 中华流行病学杂志

<p><b>OBJECTIVE</b>Using the indirect economic burden of stroke in a rural population to develop rational allocation of future health resources, in Hanzhong area.</p><p><b>METHODS</b>Cluster sampling which involved 53 natural villages with a total number of 75,000 people selected from the 'stroke monitoring base' of rural population was adopted in this study in the Hanzhong area. All of the 164 stroke cases were studied through a self-designed questionnaire. In calculating disability-adjusted life years (DALYs), fixed value was used in accordance with the value of GBD. The disability assessment was simplified in DALYs calculation and modified Barthel's ADL was used in disability assessment of stroke patients. In indirect economic burden analysis, the human capital method combined with DALYs was adopted with the formula as: indirect economic burden = GNP per capita x DALYs x productivity weight.</p><p><b>RESULTS</b>The total DALYs were 598.88, with an average DALY of stroke as 3.65 per case. The total indirect economic burden of stroke patients in rural areas was 1,993,977.8 RMB and the average of indirect economic burden of stroke was 12,158.4 RMB per case with the largest seen in the 45-59 age group, accounted for 74.4%.</p><p><b>CONCLUSION</b>In our study, the use of method in combining the human capital with DALYs was the first time being adopted in calculation of the indirect economic burden of stroke in rural population in China. The burden seemed to be much lower than literature cited from other countries. It was reasonable to evaluate indirect economic burden of stroke using method in integrating DALYs with human capital, but it was difficult to calculate the DALYs.</p>

A retrospective study on the survival rate and risk factors of mortality among 617 inpatients with ischemic stroke / 中华流行病学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to describe survival status and risk factors of mortality on inpatients with ischemic stroke.</p><p><b>METHODS</b>617 patients with continuous ischemic stroke cases were collected from January 2002 to June 2005 retrospectively in the Department of Neurology, Xijing Hospital, Fourth Military Medical University. In order to perceive relevant information on survival and the cause of death. All patients were followed through phone calls or mailing. The follow-up program was completed in January 2006. Kaplan-Meier methods were used for survival description. Monovariant and multivariant Cox's proportional hazard regression model were used to analyze prognostic factors on mortality.</p><p><b>RESULTS</b>The longest time in the follow-up program was 47 months with 59 dropped-out cases, making the dropout rate as 9.5%. Of these patients, 80 cases died during the period of study(60 for ischemic stroke,3 for cerebral hemorrhage, 10 for cardiac disease, 7 for other cause). The median survival time was 42. 16 months. The survival rates of one-year, two-year and three-year period were 91.9%, 89.4% and 85.3%, respectively. Monovariant and multivariant Cox's proportional hazard regression model showed that the risk factors associated with mortality were old age (RR = 1.043, 95% CI: 1.013-1.074), lower Glasgow scores (RR = 0.855, 95% CI: 0.742-0.985) ,poor conscious levels(RR = 4.085, 95% CI: 2.128-7.844) and having complication (RR = 1.765, 95% CI: 1.108-2.812).</p><p><b>CONCLUSION</b>The results of this study suggested that the risk factors were old age, lower Glasgow scores, poor conscious levels and having complication on mortality of ischemic stroke.</p>