Effect of DOT1L gene silence on proliferation of acute monocytic leukemia cell line THP-1 / 中国实验血液学杂志

This study was aimed to investigate the influence of short hairpin RNA (shRNA) on proliferation of human leukemia cell line THP-1. The shRNA targeting the site 732-752 of DOT1L mRNA was designed and chemically synthesized, then a single-vector lentiviral, tet-inducible shRNA-DOT1L system (Plko-Tet-On) was generated. Thereafter, the THP-1 cells with lentivirus were infected to create stable cell line with regulatable shRNA expression. The expression of DOT1L in the THP-1 cell line was assayed by RT-PCR. Effect of shRNA-DOT1L on the proliferation of THP-1 cells was detected with MTT method,and the change of colony forming potential of THP-1 cells was analyzed by colony forming unit test. Cell cycle distribution was tested by flow cytometry. The results indicated that the expression of DOT1L was statistically lower than that in the control groups. The proliferation and colony forming capacity of THP-1 cells were significantly inhibited. The percentage of cells at G0/G1 phase increased in THP-1/shRNA cells treated with Dox while the percentage of cells at S phase significantly decreased as compared with that in the control group. It is concluded that the shRNA targeting DOT1L can effectively inhibit the proliferation of acute monocytic leukemia cell line THP-1.

Inhibition of NHE1 down-regulates IL-8 expression and enhances p38 phosphorylation / 中国实验血液学杂志

This study was purposed to explore the changes of possible angiogenetic factors other than VEGF after inhibition of NHE1 and their related mechanisms. The K562 cells were treated by NHE1 specific inhibitor cariporide, the angiogenesis factors after inhibition of NHE1 were screened by using protein chip, the IL-8 expression level after cariporide treatment was detected by real-time quantitative PCR; the K562 cells with stable interference of NHE1 were constructed, the IL-8 expression level after interference of NHE1 was detected by real-time quantitative PCR; the p38 phosphorylation level in K562 cells treated with cariporide was detected by Western blot. After treatment of K562 cells with p38 inhibitor SB203580, the IL-8 expression level was decreased by real-time quantitative PCR. The results of protein chip showed that IL-8 expression decreased after cariporide treatment. Real-time quantitative PCR confirmed this inhibitory effect. The p38 phosphorylation level increased after cariporide treatment. The down-regulation of IL-8 expression induced by cariporide treatment was partially restored after K562 cells were treated with p38 inhibitor SB203580. It is concluded that the inhibition of NHE1 can inhibit IL-8 expression through up-regulation of p38 phosphorylation.

Severe coronary artery disease in Chinese patients with abdominal aortic aneurysm: prevalence and impact on operative mortality / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Little is known about the prognosis of coronary artery disease (CAD) in Chinese patients with abdominal aortic aneurysm (AAA). The aim of this study was to evaluate the predictors of in-hospital all-cause mortality of severe CAD in Chinese patients who were hospitalized for AAAs.</p><p><b>METHODS</b>From January 2003 to August 2009, 368 patients were operated on for AAAs. The clinical characteristics were retrospectively collected. The primary outcome was the in-hospital all-cause mortality. The clinical risk factors were subjected to a multivariate analysis to determine the predictors of in-hospital all-cause mortality.</p><p><b>RESULTS</b>During their hospitalization, 23% (85/368) of the patients underwent coronary angiography, which revealed significant lesions in 93% (79/85) of the patients. In 25 cases, coronary artery bypass grafting (CABG) was performed before the AAA repair and in 16 cases of percutaneous coronary intervention (PCI) was performed. Ten patients with AAA alone died before discharge, and eight patients diagnosed with AAA combined with CAD died. There was no statistical difference in the postoperative death between the two groups. The logistic analysis showed that age > 70 years and CAD (vessels ≥ 2) were the significant factors in predicting the adverse clinical outcome.</p><p><b>CONCLUSIONS</b>The prevalence of severe CAD in Chinese patients with AAAs seemed lower than those that were reported. Myocardial evaluation and subsequent revascularization before AAA surgery could improve the clinical outcome for these patients who have severe CAD.</p>

A community-based study on relations between metabolic syndrome and carotid atherosclerosis in a middle-aged population / 中华流行病学杂志

Objective To explore the association between metabolic syndromes (MS) and carotid atherosclerosis and to estimate the predictive effects of MS under 3 different definitions.Methods A cross-sectional study was conducted in 2 community-based populations in Beijing,in 2008.1266 subjects (598 men,668 women),aged 45-69,were included in the analyses.MS was defined by the criteria of International Diabetes Federation (IDF),the revised NCEPATPm (ATP Ⅲ-R) and "The Guidelines of Dyslipidemia Control for Chinese Adult" ( "Guidelines" ) in 2007.Results The prevalence rates of MS by the 3 criteria were 39.0%,43.3% and 30.9% respectively.The Kappa value for the measure of the agreement between each pair of the 3 definitions were 0.911,0.719 and 0.730 respectively.The intima-media thickness in common carotid artery (CCA-IMT) was significantly higher (P＜0.001) in all MS groups than in non-MS groups,diagnosed with the 3 criteria independent of age,gender,LDL-C,and current smoking status.After adjustment of age,gender,LDL-C,and current smoking status,the classification of MS significantly increased the risk of prevalence of carotid atberosclerotic plaques,compared to the non-MS group.OR value were 1.499 (95% CI:1.157-1.942) for IDF,1.696 (95% CI:1.314-2.189) for NCEP-R,1.763 (95% CI:1.344-2.312) for "Guideline" respectively.Conclusion Our research findings indicated that,when MS were defined with the 3 definitions,prediction on the risk of sub-clinical atberosclerosis would work beyond some of the conventional cardiovascular risk factors such as smoking,LDL-C.There might exist some differences in gender issue on the strength of association between MS when diagnosed by different criteria and carotid plaque.

The changes of expressions of the endoplasmic reticulum chaperones GRP78, GRP94, and caspase-12 following cerebral ischemia in rats / 中国脑血管病杂志

Objective: To observe the expressions of glucose-regulated protein (GRP) 78, GRP94 and caspase-12 related with endoplasmic reticulum in the striatum during focal cerebral ischemia in rats, so as to investigate the possible effect of endoplasmic reticulum molecular chaperones and apoptosis factor in the ischemic process. Methods: Sixty rats were equally randomized into 2 groups (n = 30): The sham operated group and the ischemic group (focal transient cerebral ischemia model was established with intraluminal occlusion of left meddle cerebral artery). The expression of GRP78, GRP94 and caspase-12 in rats striatum was detected by immunohistochemistry staining and semiquantitative RT-PCR at the different time points (6, 12 and 24 h after ischemia). Results: Immunohistochemistry staining and RT-PCR results illustrated that the mRNAs and protein expressions of GRP78 and GRP94 in ischemic group were lower than that in sham operated group at each time point (P < 0.05). The expression of the GRP78 and GRP94 were highest at 12 h from 6 h to 24 h (P < 0.05) in ischemic group. The expression of caspase-12 in ischemic group upgrade at 6 h and were highest at 12 h and decreased at 24 h. There was no expression in sham operated group. Conclusion: Endoplasmic reticulum-self regulated system was started through the increased expressions of GRP78 and GRP94 after striatum ischemia in rats, so as to attenuated cerebral ischemic injury. Endoplasmic reticulum-self regulated function loss because of severe cerebral ischemia. Initiation of caspase-12 apoptosis pathway is possible one of the mechanisms that priming endoplasmic reticulum apoptosis pathway in cerebral ischemic injury.

The protective mechanism of Naoxintong on cerebral ischemic injury in rats / 第二军医大学学报

Objective: To observe the influence of Naoxintong on expression of glucose-regulated protein 78(GRP78) and GRP94 in the striatum of rats with focal cerebral ischemic injury, so as to investigate their possible protective mechanism on cerebral ischemic injury. Methods: Ninety rats were equally randomized into 3 groups(n=30): sham operated group, ischemic group (Focal transient cerebral ischemia model was established with intraluminal occlusion of left middle cerebral artery) and Naoxintong pre-treated group (Treated with Naoxintong 5 d before ischemic injury). The expression of GRP78, GRP94 in rats striatum was detected by histological method, immunohistochemistry staining and semiquantitative RT-PCR at the different time points(6,12 and 24 h after ischemic treatment). Results: The focal transient cerebral ischemia model was successfully established in rats. Histological results showed that the degree of focal cerebral ischemic injury in Naoxintong pre-treated group was significantly lower than that in ischemic group. Immunohistochemistry staining and RT-PCR results illustrated that the expression of GRP78 and GRP94 in ischemic group was lower than that in sham operated group at each time point after ischemic treatment (P<0.01). The expression of GRP78 and GRP94 in ischemic group and Naoxintong pre-treated group were the lowest at 6 h and the highest at 12 h, and the expression at 24 h was lower than that at 12 h but higher than that at 6 h. Compared with the ischemic group, the expression of GRP78 and GRP94 was higher in Naoxintong pre-treated group at the same time point (P<0.05,P<0.01). Conclusion: The expression of GRP78 and GRP94 is upregulated within 12 h of cerebral ischemic injury in rats. Naoxintong can promote the expression of GRP78 and GRP94 in ischemic cerebral areas, protect the function of endoplasmic reticulum and relieve the cerebral injury caused by ischemia.