Cryopreservation of in vitro-grown shoot-tips of <Emphasis Type="Italic">Rabdosia rubescens</Emphasis> by encapsulation-dehydration and evaluation of their genetic stability

Shoot-tips of Rabdosia rubescens, excised from in vitro-grown proliferating shoots that were cold-hardened at 5°C for 3 weeks, were encapsulated in alginate beads. Subsequently, these were precultured in a mixture of 0.4 M sucrose and 2 M glycerol for 1 h and then desiccated with silica-gel to about 21% water content prior to freezing in liquid nitrogen. After thawing, about 85% of cryopreserved shoot-tips grew into true-to-type shoots and with enhanced rooting capacity. Eight single-bud sibling lines were used to assess genetic stability of these encapsulated shoot-tips. When the relative DNA content was measured by flow cytometry (FCM), no changes were observed between controls and cryopreserved shoots. Using a sequence-related amplified polymorphism (SRAP) assay, it was observed that seven out of eight cryopreserved lines showed identical banding patterns; while the eighth line displayed an absent band, amounting to a low variance rate of 0.01%. These findings suggested that it was necessary to monitor the genetic stability of recovered cryopreserved R. rubescens shoots.     

Mitogenomes provide new insights into the evolutionary history of Prodiamesinae (Diptera: Chironomidae)

Evolutionary analysis of Prodiamesinae has long been impeded by lack of information, and its phylogenetic relationship with Orthocladiinae remains questionable. Here, ten complete mitochondrial genomes (mitogenomes) of Orthocladiinae sensu lato were newly sequenced, including three Prodiamesinae species and seven Orthocladiinae species. Coupled with published mitogenomes, a total of 12 mitogenomes of Orthocladiinae sensu lato were selected for a comparative mitogenomic analysis and phylogenetic reconstruction. Mitogenomes of Orthocladiinae sensu lato are conserved in structure, and all genes arrange the same gene order as the ancestral insect mitogenome. Nucleotide composition is highly biased, and the control region displayed the highest A + T content. All protein-coding genes are under purifying selection, and the ATP8 evolves at the fastest rate. In addition, the mitogenomes of Orthocladiinae sensu lato are highly conserved, and they are practically useful for phylogenetic inference, suggesting a re-classification of Orthocladiinae by sinking Prodiamesinae as a subgroup of Orthocladiinae.     

血流阻断的缺血预处理技术在肝癌切除术中的临床应用价值

目的:探讨血流阻断的缺血预处理技术在肝癌切除术中的临床应用价值。方法:选取2010年4月至2013年7月我院收治的96名原发性肝癌并采用肝脏部分切除术进行治疗的患者,将患者随机分为观察组和对照组,每组各48例,观察组在肝脏部分切除术阻断肝门血流前先分别给予1个5 min缺血和再灌注的处理。对照组不采取任何干预措施。术前、术后1天、3天、7天时分别进行生化检查,并于术前及术后1h对Fas-mRNA表达、Caspasc-3活性及AI进行测定,观察记录患者术后的并发症情况、手术时间、术中出血量以及住院时间。结果:术后1天、3天、7天时两组间的AST、ALT、TBIL等生化指标的含量情况相比,观察组均显著优于对照组(P0.05);术后l d,两组患者ALB均有不同程度的降低,对照组低于观察组(P0.05);术后住院时间观察组为13.28±3.85天,对照组为19.48±4.92天,观察组明显低于对照组(P0.05);术后1h,两组患者的Fas-mRNA表达、Caspasc-3活性相比于阻断前均显著提高,但观察组提高幅度明显低于对照组,差异有统计学意义(P0.05);两组阻断前均未见肝细胞凋亡,术后l h时,两组组均可见肝细胞凋亡,且对照组明显高于观察组组(P0.05)。结论:血流阻断的缺血预处理技术具有操作简便、副作用小的重要特点,应用于肝癌切除术中在保护肝功能方面具有显著的优势。     

诱导神经再生治疗阿尔茨海默病的机制研究进展

阿尔茨海默病(Alzheimer's disease,AD)是一种以进行性认知障碍和记忆减退为主要特征的中枢神经退行性疾病,已经成为老年医学中最棘手的、亟待解决的问题之一. AD的病理机制仍不清楚,尚无特效治疗药物.目前,探索AD神经再生逐渐成为研究的热点领域,通过诱导神经再生可以有效地改善AD的症状.研究表明,运用药物、物理刺激或干细胞移植方法,可以提高大脑成体神经再生,是延缓AD的病理症状和认知障碍的有效治疗策略.本文综述诱导神经再生的方法及其治疗AD的作用机制,为神经再生治疗实施提供理论依据.     

基于野外观测检验动物廊道选址模型的准确性——以白头叶猴为例

廊道构建是减少栖息地破碎化负面影响的重要策略之一。目前，已经有许多模型用于动物廊道的选址，而"选址模型是否能准确预测动物迁移的实际发生位置"一直是保护生物学最为关注的问题。最小成本路径模型（LCP）和条件最小成本廊道模型（CMTC）是两种较为常用的廊道选址模型。以白头叶猴（Trachypithecus leucocephalus）为目标物种，分别运用LCP和CMTC模拟生成白头叶猴迁移廊道，将模拟结果与野外观测廊道进行对比，检验两种方法的准确性。结果表明：与野外观测实际廊道相比，LCP模型模拟结果的完全准确率为46.7%，部分准确率为20%，完全不准确率为33.3%；CMTC模型模拟结果的完全准确率为26.7%，其余73.3%为部分准确，无完全不准确的结果；总体上看，CMTC廊道的准确率较LCP高，因而CMTC模型模拟白头叶猴实际迁移廊道位置的准确性优于LCP模型。输入"源"要素类型、阻力面栅格尺度设定、栖息地土地利用类型变化以及动物迁移行为复杂性4个因素是影响该模拟结果准确性的主要原因。     

一种新型亮氨酸5-羟化酶NmLEH的异源表达、纯化及酶学性质分析 *

羟基化氨基酸是一种新型氨基酸衍生物,可广泛用作化工材料的前体物及医药合成的中间体。将来源于Nostoc minutum的新型L-亮氨酸5-羟化酶 (NmLEH) 通过重组质粒在大肠杆菌中异源表达。结果表明,在BL21(DE3) 宿主细胞中,诱导温度为25℃,IPTG诱导浓度为0.5mmol/L,诱导10h时,蛋白质表达量最高 (0.45mg/ml);通过Ni-亲和层析和凝胶过滤层析两步分离纯化获得了高度纯化的重组NmLEH蛋白;对NmLEH的酶学性质进行了表征,该酶的最适反应温度为25℃,最适pH 为7.5,在pH 7.0~9.0较为稳定,最适底物为亮氨酸和甲硫氨酸;同源序列分析表明NmLEH属于亚铁和α-酮戊二酸依赖性双加氧酶家族[Fe(II)/αKG-Dos],并预测了该酶的保守催化活性位点(H150、D152、H236);通过同源建模得到了该蛋白质的模拟结构,分析了该蛋白质催化活性中心的形成机制。     

引进马铃薯种质资源抗旱性评价

马铃薯属于干旱敏感型作物,当前生产上的马铃薯品种多数不耐旱,中国马铃薯抗旱育种进程又受到遗传背景狭窄的制约。引进外来种质资源,拓宽我国马铃薯遗传背景,加快选育抗旱品种是马铃薯应对干旱的关键策略。2016年和2017年,在常规滴灌和雨养条件下,利用增广设计方法,以生产上常用的5个马铃薯品种为对照,对来自国际马铃薯中心的315份高代品系和中国已有的3个品种进行抗旱性评价。通过AMMI模型和GGE模型分析基因型、环境及二者互作对产量的影响,并结合抗旱指数筛选抗旱性稳定且产量高的材料。从整体上看,在雨养条件下,两年马铃薯平均产量差异较小,但是变异系数较大,常规滴灌条件下正好相反。马铃薯产量受基因型、环境及其交互作用的显著影响,其变异平方和分别占总处理平方和的43.39%、39.36%和17.26%;C93和YS902两年的抗旱指数均高于对照品种,稳产性好,C48虽然抗旱指数相对较低,但是高产和稳产性高于所有材料。筛选出来的材料不仅可以作为抗旱育种亲本,还可以通过进一步研究其抗旱机制,为抗旱育种提供理论支持。