目的 采用RNA干扰技术沉默CCS（copper chaperone for SOD1）基因，构建相关小干扰RNA（siRNA），探索出针对CCS的高效siRNA序列。 方法 合成用于人脐静脉内皮细胞（HUVEC）细胞中沉默CCS基因的siRNA。应用脂质体转染的方法在HUVEC细胞中对CCS基因进行RNA沉默。蛋白免疫印迹Western blotting检测沉默前后CCS蛋白表达变化的情况，甲基四唑蓝法MTT检测转染前后细胞活力。最后用单因素方差分析对数据进行统计学分析，以确定有效的siRNA序列。 结果 转染前后细胞形态无肉眼可见变化，转染后细胞活力分别为98.5%和98.8%。CCS蛋白沉默率分别为63.7%和61.4%。 结论 采用siCCS-2和siCCS-3序列转染条件对HUVEC细胞活力损伤小，CCS沉默效率高，实验条件稳定，重复性好。为我们继续研究沉默CCS后抑制血管内皮细胞的生长增殖、血管形成提供了稳定的实验基础。
ObjectiveThis study examined the pattern of adjunctive antidepressant use in schizophrenia patients and its demographic and clinical correlates in a nationwide survey in China. MethodsFourteen thousand and thirteen patients in 45 Chinese psychiatric hospitals or centers were interviewed (4,486 in 2002, 5,288 in 2006, and 4,239 in 2012). Patients' sociodemographic and clinical characteristics were recorded using a standardized protocol and data collection procedure. Chi-square test, independent-samples t test, Mann-Whitney U test, and multiple logistic regression analysis were used in data analyses. ResultsAntidepressant use was found in 5.2% of the study population with 4.6% in 2002, 4.3% in 2006, and 6.9% in 2012, respectively. A significant increase in use from 2006 to 2012 was found (p<.001). Multiple logistic regression analyses in the whole population revealed that patients receiving adjunctive antidepressants were more likely to be outpatients in tertiary referral centers (level-III hospitals) and who had an earlier age of onset, less severe global illness, but more depressive symptoms. They were less likely to receive first-generation antipsychotics but more likely to receive benzodiazepines (R-2=0.255, p<.001). ConclusionsDespite an increasing trend, the frequency of antidepressant use in schizophrenia in China was considerably lower than in Western countries. The benefits and risks associated with concomitant use of antidepressants in schizophrenia need to be studied further.
alpha-Calcitonin gene-related peptide (alpha-CGRP) plays a significant pathophysiological role in bone development, metabolism and remodeling around dental implants. However, the half-life of alpha-CGRP in plasma is only 10 min, which affects its long-time application and an alternative approach should be developed to deliver alpha-CGRP over long periods of time. The aim of this study is to investigate whether a lentiviral alpha-CGRP overexpression vector system can express this target-gene longer at peri-implant sites, thus enhancing osseointegration. Animals were divided to the following groups: alpha-CGRP(-/-), alpha-CGRP(-/-) with lentivirus transfection and alpha-CGRP(+/+) mice. MS Spectrum imaging observations identified the successful transfection of alpha-CGRP around experimental implants inserted in the femurs at 5 days after injection. Histomorphometrical analysis indicated an increase of bone-implant contact (BIC) at 1-month healing in the transfection group. Moreover, real-time RT-PCR and western blot results of bone-related markers Runx2, Osterix, and BSP levels elevated in lentivirus-transfected mice at 21 days, compared to the untreated alpha-CGRP(-/-) mice. There was no significant difference between the transfection group and alpha-CGRP(+/+) group. Further alpha-CGRP protein detection confirmed the persistent expression of this transgene at 21 days post-operatively. These results suggest that this lentiviral vector system expresses alpha-CGRP in an effective, appropriate and sustained manner, which might have a potential application in enhancing titanium implant osseointegration. (C) 2016 Elsevier Inc. All rights reserved.