结晶型硫化镍诱发细胞恶性转化过程中基因组总体DNA甲基化的改变

Objective To observe the effect of crystalline NiS on genome DNA methylation profile in in vitro cultured cells.Methods 16HBE Cells were treated with crystalline NiS at 0.25,0.50,1.00 and 2.00 μg/cm2 for 24 h and three times at total.DAC treatment was given at 3 μmol/L for 72 h.5-mC immunofluorescence and SssI emthyltrasferase assay methods were applied to investigate if the hypomethylation of genome DNA involved.Results The results of 5-mC immunofluorescence showed that the fluorescence intensity of NiS-treated cells were decreased in some degree, and transformed cells were decreased dramatically.By the SssI methylase assay, an average of (81.9 ± 7.3 )% methylated CpG were found in negative control cells.By contrast, ( 77.9 ± 6.2) %, ( 75.3 ± 6.8 ) %, ( 59.5 ± 4.9 ) %, ( 67.4 ±5.1 ) % methylated CpG were observed in cells treated with NiS for three times at dosage of 0.25,0.50,1.00and 2.00 μg/cm2 which were abbreviated as NiS0.25, NiS0.50, NiS1.00, NiS2.00 respectively.The ANOVA analysis results showed that there was a significant difference in the 5 groups above ( F = 124.95,P ＜0.01 ).The results of Dunnett-t test showed that the methylated CpG of both group NiS1.00 and NiS2.00were significantly decreased compared with the negative control group(t values were 7.64,4.89 respectively,P ＜0.01 ).For methylated CpG, (46.2 ±4.1 ) % and (43.6% ±4.3)% were observed in NiS-transformed cells (NSTC1 and NSTC2) which were dramatically decreased compared with the negative control group(t values were 12.79,13.56 respectively, P ＜ 0.01 ).Conclusion Genomic DNA methylation levels were decreased during NiS induced malignant transformation.     

紫外线照射所致的DNA损伤对连接转化的影响

目的探讨DNA经不同波长紫外线照射与经紫外线照射不同时间所致损伤对基因库构建中DNA连接转化效率的影响。方法经改造的质粒DNA使用SfiI酶切过夜,1%琼脂糖凝胶电泳分离,在不同波长不同时间紫外线照射的条件下回收目的片断,并进行目的片断的连接转化,计数菌落数,计算转化效率。结果经302nm波长紫外线照射后,其连接转化的菌落数极少(60个/ngDNA),365nm波长紫外线照射条件下回收的DNA仍可被有效连接转化。在365nm波长紫外线照射下,与未经照射进行比较,当照射时间在30min,其连接转化成功率低于50%,而照射5min、15min的连接转化成功率仍高于70%,可满足大部分分子生物学实验的需要。回收的DNA连接转化的最大效率可超过20000个菌落/1ngDNA。结论在构建大容量基因库回收目的片断时,应避免紫外线的照射;在需要时,应选择在365nm波长紫外线照射条件下进行,且照射时间应尽量缩短,以不超过15min为宜。     

Comparison of Recurrence Rates with Contour-Loop Excision of the Transformation Zone (C-LETZ) and Large Loop Excision of the Transformation Zone (LLETZ) for CIN

Aim: To compare recurrence rates of large loop excision of the transformation zone (LLETZ) with those of contour-loop excision of the transformation zone (C-LETZ) in the management of cervical intraepithelial neoplasia (CIN). Materials and Methods: The medical records of 177 patients treated consecutively by LLETZ and C-LETZ for CIN at Rajavithi Hospital between 2006 and 2009 were retrospectively reviewed. Results: Of the 87 women in the C-LETZ group, 2 cases (2.30%) had recurrence compared with 13 cases (14.4%) of the 90 women in the LLETZ group, the higher recurrence rate in the latter being statistically significant (p<0.05). Median times of follow up in the C-LETZ and LLETZ groups were 12 months and 14 months respectively (p>0.05). The C-LETZ group showed less intraoperative bleeding compared to the LLETZ group, but the rate of achievement of single specimens and positive margins were similar in the two groups. Conclusions: The present study demonstrated the superiority of C-LETZ over LLETZ in terms of efficacy; C-LLETZ is associated with a lower recurrence rate and also carries a smaller risk of intraoperative bleeding than LLETZ. The rotatingtechnique still has a potential role in treating precancerous lesions of the cervix.     

卵巢子宫内膜异位症恶变与卵巢癌的相关性研究及预后分析

目的:探讨不同原因所致卵巢癌,尤其是是否由卵巢子宫内膜异位症(EMT)恶变而来的卵巢癌的临床病理及预后特征。方法:纳入2006年1月至2011年8月在四川大学华西第二医院病理检查确诊的有随访资料(术后随访时间2~89月)的EMT恶变者所致原发性卵巢癌患者49例为EMT恶变组,其病理类型为透明细胞癌、子宫内膜样腺癌及混合性腺癌;另选择非EMT恶变所致卵巢癌并与EMT恶变者具有相同组织学类型的卵巢癌患者188例为非EMT恶变组,比较两组卵巢癌的临床病理特征及预后特征。结果:1与非EMT恶变组比较,EMT恶变组在40岁人群中所占比例较大、CA125更多表现为正常而非升高、FIGO分期早期病例比例更多、透明细胞癌及子宫内膜样腺癌这两种病理类型的比例较高,两组差异有统计学意义(P0.05)。2单因素预后分析:EMT恶变所致透明细胞癌的生存率明显高于非EMT恶变所致透明细胞癌(P=0.010),是否由EMT恶变对子宫内膜样腺癌及混合性腺癌的预后影响不确切(P0.05)。3多因素COX回归分析:EMT恶变、手术满意度及FIGO分期是影响预后的独立危险因素。结论:EMT恶变所致卵巢癌病理类型主要包括透明细胞癌和子宫内膜样腺癌,EMT恶变是卵巢癌预后的独立影响因素,EMT恶变患者年龄相对较年轻、FIGO分期相对较早,尤其对透明细胞癌而言,EMT恶变的患者可能拥有更好的预后。     

基于小波和卡尔曼平滑的事件相关电位单次提取

本研究提出一种事件相关电位单次提取方法,可有效减少实验次数,并可探索实验之间ERP的变异性。此方法基于小波和卡尔曼平滑,首先利用小波变换考察ERP平均信号的时频特性,根据ERP不同分量出现的时间位置,在不同尺度上选取特定的单次实验ERP小波系数构成观测向量,其为真实ERP小波系数状态向量与噪声之和,然后对观测向量进行卡尔曼平滑,最后对卡尔曼平滑后的小波系数进行小波重构,得到单次提取的ERP信号。仿真实验表明,基于小波和卡尔曼平滑的方法不仅信噪比提高约16~18 dB,优于30次叠加平均、简单小波方法和基于高斯基函数的卡尔曼滤波方法,还可以跟踪ERP的幅度趋势变异性。与基于高斯基函数的卡尔曼滤波方法相比,所提方法降低了计算量。真实脑电ERP提取实验表明本方法较好地从单次记录中提取出了事件相关电位,并可解释ERP因适应和应激引起的趋势变异性。     

皮肤药物不良反应的研究进展

药物不良反应十分常见,而皮肤是其重要的靶器官之一.皮肤药物不良反应(cutaneous adverse drug reaction,CADR)的发病机制复杂,临床表现多样,病因诊断较为困难.现概述CADR的特殊临床表现、发生机制和诊断方法.一、药物皮肤不良反应的临床表现1.痤疮样疹或脓疱型药疹:痤疮样疹可由碘剂、溴剂、促肾上腺皮质激素、糖皮质激素、异烟肼、雄激素、锂剂、放线菌素D和苯妥英引起[1].     

糖尿病肾病中医药诊治进展

糖尿病肾病是糖尿病患者的主要微血管病变之一,亦是导致终末期肾脏病的重要原因之一。中医药诊治糖尿病肾病有多种理论学说,从不同角度阐释了糖尿病肾病的病机,其中大部分医家认为气阴两虚,肾络瘀阻为主要病因病机。文章将糖尿病肾病中医药诊治进展进行了总结。     

Effect of body position on the pathogenesis of asymmetric primary open angle glaucoma

AIM: To explore the pathogenesis of asymmetric primary open angle glaucoma (POAG) in both eyes by comparing the intraocular pressure (IOP) of patients who sleep in different positions and to investigate the relationship between IOP variations and sleep positions. METHODS: One hundred and thirty-one patients with asymmetric POAG and forty-six healthy volunteers were enrolled. All participants completed a questionnaire that provided information about their sleep laterality. The cup disc ratio (C/D) and visual field defect established binocular asymmetry. The IOP of both eyes was measured using iCare parameters after the patients were asked to change body position. The “worse” and “better” eyes were identified according to the diagnosis, whereas the “dependent” and “independent” eyes were defined according to the lateral position. RESULTS: No significant difference in sleep laterality was observed between healthy people and patients with POAG (F=3.195, P=0.362). Among the enrolled patients, the IOP of the dependent eye was always greater than that of the independent eye in the lateral position (P<0.05). In the patients with binocular asymmetric POAG, the questionnaire clearly showed that 85.7% of left side preferences were found their left eyes to be the worse eyes and the right eyes of 71.4% patients with a right side preference were the more serious. When the asymmetric C/D ratio was greater than or equal to 0.2, the worse eye of patients with POAG and a preferred sleeping position was the dependent eye (χ2=16.762, P=0.001). CONCLUSION: A higher IOP was measured in the dependent eye in the lateral position. The long-term tendency to choose a lateral sleeping position might lead the dependent eye to manifest more severe symptoms than the independent eye. Thus, the lateral sleeping position might be one cause of asymmetric POAG.     

药用植物中转基因元件筛查策略研究

通过对已有药用植物转基因研究文献进行总结分析,为药用植物和中药材转基因监管提供有效策略。采用关键词检索法对中国知网和SCI两大数据库进行查询,建立48种常用药用植物转基因研究中常用表达元件的数据库,分析转基因元件使用情况,并总结外源转基因的筛查策略。获得1993年4月至2016年5月的转基因研究文献281篇,包括中文文献230篇,英文文献51篇。40.4%的中文文献和54.9%的英文文献研究目的为优化药用植物遗传转化体系。常用启动子包括P-35S、P-Ubi、P-GPD和P-act,其中P-35S的使用频率最高,为68.7%。常用标记基因包括NPTII、HPT、Gent、Bar和aad A,其中NPTII的使用频率最高,为37.4%。常用报告基因包括GUS和GFP,GUS使用频率达到35.2%。常用终止子包括T-NOS、T-35S和T-OCS,其中T-NOS的使用频率最高,为58%。对各元件进行配组分析,发现"P-35S+T-NOS+NPTII+GUS"组合筛选成功率为86.1%,在此基础上加入有一定使用频率的HPT、Bar和GFP,对药用植物转基因元件的筛查率达到91.5%。对于使用同源或内源启动子、标记基因和终止子的研究,可基于T-DNA边界序列进行转基因检测。