Comparative Anatomy of Chromosomal Domains with Imprinted and Non-Imprinted Allele-Specific DNA Methylation

Allele-specific DNA methylation (ASM) is well studied in imprinted domains, but this type of epigenetic asymmetry is actually found more commonly at non-imprinted loci, where the ASM is dictated not by parent-of-origin but instead by the local haplotype. We identified loci with strong ASM in human tissues from methylation-sensitive SNP array data. Two index regions (bisulfite PCR amplicons), one between the C3orf27 and RPN1 genes in chromosome band 3q21 and the other near the VTRNA2-1 vault RNA in band 5q31, proved to be new examples of imprinted DMRs (maternal alleles methylated) while a third, between STEAP3 and C2orf76 in chromosome band 2q14, showed non-imprinted haplotype-dependent ASM. Using long-read bisulfite sequencing (bis-seq) in 8 human tissues we found that in all 3 domains the ASM is restricted to single differentially methylated regions (DMRs), each less than 2kb. The ASM in the C3orf27-RPN1 intergenic region was placenta-specific and associated with allele-specific expression of a long non-coding RNA. Strikingly, the discrete DMRs in all 3 regions overlap with binding sites for the insulator protein CTCF, which we found selectively bound to the unmethylated allele of the STEAP3-C2orf76 DMR. Methylation mapping in two additional genes with non-imprinted haplotype-dependent ASM, ELK3 and CYP2A7, showed that the CYP2A7 DMR also overlaps a CTCF site. Thus, two features of imprinted domains, highly localized DMRs and allele-specific insulator occupancy by CTCF, can also be found in chromosomal domains with non-imprinted ASM. Arguing for biological importance, our analysis of published whole genome bis-seq data from hES cells revealed multiple genome-wide association study (GWAS) peaks near CTCF binding sites with ASM.     

脐带问充质干细胞联合表皮生长因子治疗III、Ⅳ期压疮的临床研究

目的：探讨脐带间充质干细胞移植联合表皮生长因子治疗III、Ⅳ期压疮的临床效果。方法：选取我院2011-2012年收治的III、Ⅳ期压疮患者143例为研究对象,随机分为对照组、实验一组和实验二组。对照组30例压疮患者行常规换药,实验一组58例压疮患者应用脐带间充质干细胞移植联合表皮生长因子治疗,实验二组55例压疮患者仅应用表皮生长因子治疗。治疗后,比较三组患者愈合时间及愈合效果。结果：实验一组和实验二组的显效时间和愈合时间均较对照组显著缩短,且实验一组的显效时间和愈合时间较实验二组更短,差异均有统计学意义（均P〈0．05）。实验一组的治愈率显著高于对照组（P〈0．05）,而与实验二组比较无统计学差异（P〉0．05）,实验二组的治愈率与对照组比较无统计学差异（P〈0．05）。实验一组的总有效率显著高于对照组和实验二组（P〈0．05）,而实验二组的总有效率与对照组比较无统计学差异（P〉0．05）。结论：脐带间充质干细胞移植联合表皮生长因子较单用表皮生长因子及常规治疗方法更好的改善III、Ⅳ期压疮创面的愈合,避免患者组织感染,提高压疮患者的生活质量。     

Association of Dyslipidemia with Renal Cell Carcinoma: A 1∶2 Matched Case-Control Study

Abnormal serum lipid profiles are associated with the risk of some cancers, but the direction and magnitude of the association with renal cell carcinoma is unclear. We explore the relationship between serum lipids and renal cell carcinoma via a matched case-control study. A 1∶2-matched case-control study design was applied, where one renal cell carcinoma patient was matched to two non-renal-cell-carcinoma residents with respect to age (±0 year) and gender. Cases (n = 248) were inpatients with a primary diagnosis of renal cell carcinoma, confirmed by pathology after operations. Controls were sampled from a community survey database matched on age and gender with cases, 2 controls for each case. Stratified Cox proportional hazard regression analysis was used to obtain hazard ratios and corresponding 95% confidence intervals of lipids level and dyslipidemia for the risk of renal cell carcinoma. Elevated serum cholesterol (p<0.001), LDL cholesterol (p<0.001), and HDL cholesterol (p = 0.003) are associated with decreased hazard of renal cell carcinoma, adjusting for obesity, smoke, hypertension and diabetes. However, risk caused by hTG showed no statistical significance (p = 0.263). This study indicates that abnormal lipid profile influences the risk of renal cell carcinoma.     

细胞色素P450酶的结构、功能与应用研究进展

细胞色素P450 (cytochrome P450,CYP)酶是广泛存在于微生物、动植物及人体中与膜结合的血红蛋白类酶,具有氧化、环氧化、羟化、去甲基化等多种生物催化活性。CYP酶在药物、类固醇、脂溶性维生素和许多其他类型化学物质的代谢中具有重要作用,其在异源物质的解毒、药物相互作用和内分泌功能等领域的研究是热点问题。本综述对CYP的结构、功能、临床应用与开发前景进行了概述,并对其最新的研究现状和发展前景进行探讨。     

不同牛分枝杆菌特异性基因PCR方法的比较

【背景】牛结核病是我国二类动物疫病,世界动物卫生组织将其列为法定报告的动物疫病。牛主要通过患病牛呼吸道分泌物和咳嗽所产生的气溶胶感染;人则主要通过食用未经高温处理的病牛的肉或奶感染。因此,经过病原学PCR检测对疑似患病牛牛奶或屠宰组织样品进行快速检验确诊,能够最大限度地减少奶牛养殖中乳品生产业的经济损失。【目的】研究并确定适宜的牛分枝杆菌PCR扩增引物及参数,为临床快速准确诊断牛结核病提供参考。【方法】对已报道的5对PCR引物,运用降落(touch down) PCR法确定适宜退火温度(Tm);运用梯度稀释的牛分枝杆菌C68001株(国内牛结核菌素生产用菌株)基因组DNA以及不同菌液含量的人工模拟临床样本(淋巴结、肺脏和牛奶),确定不同引物PCR方法的敏感性;同时以6种常见牛感染菌(牛种布鲁氏菌2308、羊种布鲁氏菌Rev.1、牛分枝杆菌C68001和AN5、禽分枝杆菌C68202、副结核分枝杆菌C68681和胞内分枝杆菌C68226)核酸样本,确定不同引物PCR方法的特异性。【结果】所有引物在53-63℃均含有目的条带,确定引物的最佳退火温度是60℃。在细菌核酸敏感性检验中,1号和3号引物的检测敏感性最高,达10-10 ng/μL;其次是2号和5号,达10-5 ng/μL。对于人工模拟感染样本,1号、3号和4号引物在淋巴结和肺脏中检测敏感性最高,其次是2号;而2号、3号、4号和5号引物对奶样检测敏感性最高。对于特异性检验,2号和5号引物特异性较好,可检测到明显的牛分枝杆菌特异性条带,对通常不引起牛结核病而只干扰免疫学诊断的禽分枝杆菌检测条带较微弱,而布鲁氏菌、副结核分枝杆菌和胞内分枝杆菌均无检测条带。【结论】2号引物及其反应参数的PCR方法敏感性、特异性良好,适合用于牛结核病的快速准确诊断。     

肠道细菌弗氏柠檬酸杆菌和产酸克雷伯氏菌对斑翅果蝇生长发育和物质代谢的影响

【目的】明确弗氏柠檬酸杆菌Citrobacter freundi和产酸克雷伯氏菌Klebsiella oxytoca两种肠道共生细菌对斑翅果蝇Drosophila suzukii生长发育和物质代谢的影响。【方法】以正常饲养条件下的斑翅果蝇、构建的斑翅果蝇无菌品系以及弗氏柠檬酸杆菌和产酸克雷伯氏菌单一共生菌感染的斑翅果蝇品系为材料,检测不同品系间斑翅果蝇的卵孵化率、3龄幼虫体重和化蛹率;测定不同斑翅果蝇品系3龄幼虫体内蛋白质、氨基酸、糖原和游离脂肪酸等代谢物的含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD)的活力。【结果】正常饲养条件下的斑翅果蝇卵孵化率、3龄幼虫体重、化蛹率及3龄幼虫体内蛋白质的含量均高于其他斑翅果蝇品系,且无菌品系中的最低。弗氏柠檬酸杆菌和产酸克雷伯氏菌感染的斑翅果蝇品系3龄幼虫中的氨基酸和糖原含量均低于斑翅果蝇无菌品系和正常品系。弗氏柠檬酸杆菌感染斑翅果蝇品系3龄幼虫体内游离脂肪酸的含量较其他品系的也降到最低。弗氏柠檬酸杆菌和产酸克雷伯氏菌感染斑翅果蝇品系3龄幼虫体内POD活力显著高于无菌品系和正常品系,而CAT活力显著低于无菌品系。【结论】斑翅果蝇肠道中无肠道共生细菌时生长发育迟缓,在食物中分别添加弗氏柠檬酸杆菌和产酸克雷伯氏菌后可一定程度上促进斑翅果蝇的发育,这与添加肠道共生菌后斑翅果蝇体内代谢物的变化密切相关。     

链霉菌质粒的复制与进化

近年来, 随着大质粒提取和检测技术的发展, 尤其是高通量DNA测序技术的应用, 使得链霉菌大的环型质粒和线型质粒的研究取得了较快的进展。相比于研究透彻的细菌Theta型复制的质粒, 链霉菌Theta型质粒在复制区的结构、复制蛋白和调控蛋白作用的分子机理等方面具有多样性和新颖性。新鉴定的许多线型质粒的中心复制区表明中心复制的起始可以靠近端粒, 一个质粒也可以有2个以上的复制区。新分离的端粒序列显示端粒“折返”不是必需的, 而形成二级结构对于端粒复制是重要的。链霉菌环型和线型质粒的测序分析显示它们之间存在亲缘关系。环型质粒可以与噬菌体共整合, 实验证明它们在一定条件下可以相互转换。这些研究结果表明, 链霉菌环型、线型质粒和噬菌体从结构到功能到进化具有多样性、新颖性和亲缘关系。