流体动力对HEK293细胞的细胞团形成及细胞生长和代谢的影响

利用HEK293细胞在悬浮培养中具有聚集成团的体外培养特性,在250mL的Bellco的搅拌培养体系中,以HEK293细胞团的粒径、细胞数、细胞活力、葡萄糖比消耗率(qglc)、乳酸比产率(qlac)和乳酸转化率(Ylacglc)为观察指标,考察HEK293细胞在搅拌速度分别设置为25、50、75和100rmin的培养条件下的细胞团形成、粒径分布以及细胞生长和代谢。HEK293细胞在搅拌速度为50rmin和75rmin培养条件下所形成细胞团的粒径大小适中、离散度小。培养7d后,HEK293细胞团的平均粒径分别为201μm和175μm,其中粒径≥225μm的细胞团所占比例均低于10%;在整个培养过程中,细胞团中的HEK293细胞活力维持在90%以上,qglc、qlac和Ylacglc等反映HEK293细胞代谢的参数保持相对恒定。实验结果提示:合适的搅拌速度所产生的流体动力既可使细胞团的粒径控制在合适的范围内,也可为细胞团中的HEK293细胞提供基本满足其正常生长和代谢需要的物质传递效率。     

卷叶黄精对植物病原菌的抗菌活性研究

采用菌丝生长速率法研究了卷叶黄精根提取物对植物病原菌的抗菌活性。结果表明:卷叶黄精对所选10种植物病原菌中的7种呈现不同程度的抗菌活性,而且各萃取物的抑菌菌种各不相同,其中正丁醇萃取物对4种病原菌、乙酸乙酯萃取物对3种病原菌有明显的抑菌活性,水浓缩物仅对1种病原菌有抗菌活性,而石油醚萃取物对供试植物病原菌没有抗菌活性;正丁醇萃取物对苹果褐腐病、玉米大斑病、棉黄萎病病原菌抑制作用较强,EC50分别为261.26、376.03和430.05μg.mL-1;乙酸乙酯萃取物对苹果腐烂病、番茄黑霉病病原菌抑制作用较强,EC50分别为386.00和865.75μg.mL-1,各样品对植物病原菌的毒力随着浓度的增高而增强;卷叶黄精对植物病原菌的抑菌活性随着作用时间的延长,均呈现不同的下降趋势。     

基于滑动窗口的原核转录起始位点计算定位方法

转录起始位点的计算定位是基因转录调控研究的重要内容,但现有方法的识别性能较低。文章作者在已有原核启动子识别算法的基础上,提出了一种基于滑动窗口的原核转录起始位点计算定位方法,通过在合理限定的定位范围内对序列进行滑动扫描,来预测转录起始位点的位置。首先根据窗口序列的交迭组分特征和启动子其它特征分别建立二次判别分类器,用其计算对应位置的似然得分,再利用转录起始位点与翻译起始位点的间隔经验分布信息对似然得分进行修正,最后依照似然得分的分布情况由阈值定位算法确定预测位置。对大肠杆菌真实序列数据的测试结果表明,该定位算法可实现对真实转录起始位点位置的有效预测,与已有算法相比,当敏感性指标同为0.85左右时,特异性指标可从0.20提高至0.65,从而使得定位准确率提高了约20个百分点。     

土壤动物对川西亚高山和高山森林凋落叶第一年不同分解时期N和P元素动态的影响

土壤动物对高寒森林凋落物养分元素动态具有重要影响, 但这种影响受控于凋落物质量及环境条件。为了解土壤动物对高寒森林凋落物不同分解时期凋落物中N和P元素动态的影响, 采用凋落物分解袋的方法, 于凋落物第一年分解的不同时期, 即冻结前期、冻结期、融化期、生长季节初期、生长季节中期和生长季节末期, 研究了3.00和0.04 mm孔径凋落物袋中川西亚高山和高山森林的代表性植物——康定柳(Salix paraplesia)、方枝柏(Sabina saltuaria)、红桦(Betula albosinensis)和岷江冷杉(Abies fargesii var. faxoniana)凋落物中的N和P元素动态特征。结果表明: 康定柳和红桦凋落物中的N元素呈现出释放—富集—释放的模式, 方枝柏、岷江冷杉凋落物中的N元素则表现为释放—富集模式; 凋落物P元素总体表现为释放模式, 但4种植物凋落物均在生长季节中期具有明显的富集过程; 从凋落物分解的第一年来看, 土壤动物明显促进了4种植物凋落物N的释放, 而抑制了P的释放; 不同时期土壤动物对凋落物中N和P释放量的影响存在显著差异, 且分别与正积温呈极显著正相关和极显著负相关关系; 相对于阔叶植物凋落物, 土壤动物对针叶植物凋落物中N和P元素动态的影响更为显著。这些结果为深入了解高寒森林生态系统土壤动物与凋落物分解等物质循环过程的相互联系具有重要意义。     

lncRNA在肿瘤中的表达及作用机制

长链非编码RNA(long non-coding RNAs,lncRNAs)是一类转录本长度大于200 nt,不具有蛋白编码功能的非编码RNA.近年来,随着高通量测序技术的发展,越来越多的功能性lncRNAs逐渐被发现,且已成为研究的热点.研究发现,lncRNAs可以作为致癌或抑癌基因参与肿瘤的发生发展.通过比对肿瘤细胞和正常细胞的表达谱显示,多种lncRNAs在不同类型肿瘤中异常表达.除了表型上的研究,目前已有部分报道深入研究了lncRNAs在肿瘤中的作用机制.例如,lncRNAs与肿瘤细胞凋亡、转移及耐药等过程密切相关.此外,在肿瘤患者的临床常规检验中检测到lncRNAs,提示其能够作为一种潜在的肿瘤诊断和预后因子. lncRNAs有望成为新型肿瘤标志物和肿瘤治疗的靶点,用于诊断、治疗、预测预后.本文将通过对lncRNAs在肿瘤中的作用及其分子机制进行综述.     

Lactase persistence in central Asia: phenotype, genotype, and evolution

The aim of the present study is to document the evolution of the lactase persistence trait in Central Asia, a geographical area that is thought to have been a region of long-term pastoralism. Several ethnic groups co-exist in this area: Indo-Iranian speakers who are traditionally agriculturist (Tajik) and Turkic speakers who used to be nomadic herders (Kazakh, Karakalpak, Kyrgyz, Turkmen). It was recently demonstrated that horse milking practice existed in the Botai culture of Kazakhstan as early as 5,500 BP ( Outram et al. 2009 ). However, the frequency of the lactase persistence trait and its genetic basis in Central Asian populations remain largely unknown. We propose here the first genotype-phenotype study of lactase persistence in Central Asia based on 183 individuals, as well as the estimation of the time of expansion of the lactase-persistence associated polymorphism. Our results show a remarkable genetic-phenotypic correlation, with the causal polymorphism being the same than in Europe (-13.910C>T, rs4988235). The lactase persistence trait is at low frequency in these populations: between 25% and 32% in the Kazakh population (traditionally herders), according to phenotype used, and between 11% and 30% in the Tajiko-Uzbek population (agriculturalists). The difference in lactase persistence between populations, even if small, is significant when using individuals concordant for both excretion of breath hydrogen and the lactose tolerance blood glucose test phenotypes (P = 0.018, 25% for Kazakh vs. 11% for Tajiko-Uzbeks), and the difference in frequency of the -13.910*T allele is almost significant (P = 0.06, 30% for Kazakhs vs. 19% for Tajiko-Uzbeks). Using the surrounding haplotype, we estimate a date of expansion of the T allele around 6,000-12,000 yrs ago, which is consistent with archaeological records for the emergence of agropastoralism and pastoralism in Central Asia.     

Evidence for a role of raffinose in stabilizing photosystem II during freeze–thaw cycles

A role of non-reducing sugars like sucrose and raffinose in the protection of plant cells against damage during freezing has been proposed for many species, but reports on physiological effects are conflicting. Non-aqueous fractionation of mesophyll cell compartments in Arabidopsis thaliana was used to show that sucrose and raffinose accumulate in plastids during low temperatures, pointing to a physiological role in protecting the photosynthetic apparatus. Comparing a previously described raffinose synthase (RS) mutant of A. thaliana with its corresponding wild type, accession Col-0, revealed that a lack of raffinose has no effect on electrolyte leakage from leaf cells after freeze–thaw cycles, supporting that raffinose is not essential for protecting the plasma membrane. However, in situ chlorophyll fluorescence showed that maximum quantum yield of PS II photochemistry (F _v/F _m) and other fluorescence parameters of cold acclimated leaves subjected to freeze–thaw cycles were significantly lower in the raffinose synthase mutant than in the corresponding wild type, indicating that raffinose is involved in stabilizing PS II of cold acclimated leaf cells against damage during freezing.     

Regulatory RNAs in Haloferax volcanii

In organisms of all three domains of life, a plethora of sRNAs (small regulatory RNAs) exists in addition to the well-known RNAs such as rRNAs, tRNAs and mRNAs. Although sRNAs have been well studied in eukaryotes and in bacteria, the sRNA population in archaea has just recently been identified and only in a few archaeal species. In the present paper, we summarize our current knowledge about sRNAs and their function in the halophilic archaeon Haloferax volcanii. Using two different experimental approaches, 111 intergenic and 38 antisense sRNAs were identified, as well as 42 tRFs (tRNA-derived fragments). Observation of differential expression under various conditions suggests that these sRNAs might be active as regulators in gene expression like their bacterial and eukaryotic counterparts. The severe phenotypes observed upon deletion and overexpression of sRNA genes revealed that sRNAs are involved in, and important for, a variety of biological functions in H. volcanii and possibly other archaea. Investigation of the Haloferax Lsm protein suggests that this protein is involved in the archaeal sRNA pathway.     

The role of leptin in striped hamsters subjected to food restriction and refeeding

Food restriction(FR) and refeeding(Re) have been suggested to impair body mass regulation and thereby making it easier to regain the lost weight and develop over-weight when FR ends. However, it is unclear if this is the case in small mammals showing seasonal forging behaviors. In the present study, energy budget, body fat and serum leptin level were measured in striped hamsters that were exposed to FR-Re. The effects of leptin on food intake, body fat and genes expressions of several hypothalamus neuropeptides were determined. Body mass, fat content and serum leptin level decreased during FR and then increased during Re. Leptin supplement significantly attenuated the increase in food intake during Re, decreased genes expressions of neuropepetide Y(NPY) and agouti-related protein(AgRP) of hypothalamus and leptin of white adipose tissue(WAT). Hormone-sensitive lipase(HSL) gene expression of WAT increased in leptin-treated hamsters that were fed ad libitum, but decreased in FR-Re hamsters. This indicates that the adaptive regulation of WAT HSL gene expression may be involved in the mobilization of fat storage during Re, which partly contributes to the resistance to FR-Re-induced overweight. Leptin may be involved in the down regulations of hypothalamus orexigenic peptides gene expression and consequently plays a crucial role in controlling food intake when FR ends.     

Exaggerated root respiration accounts for growth retardation in a starchless mutant of Arabidopsis thaliana

The knock‐out mutation of plastidial phosphoglucomutase (pgm) causes a starchless phenotype in Arabidopsis thaliana, and results in a severe growth reduction of plants cultivated under diurnal conditions. It has been speculated that high soluble sugar levels accumulating during the light phase in leaf mesophyll might cause a reduction of photosynthetic activity or that shortage of reduced carbon during the night is the reason for the slow biomass gain of pgm. Separate simultaneous measurements of leaf net photosynthesis and root respiration demonstrate that photosynthetic activity per unit fresh weight is not reduced in pgm, whereas root respiration is strongly elevated. Comparison with a mutant defective in the dominating vacuolar invertase (AtβFruct4) revealed that high sucrose concentration in the cytosol, but not in the vacuole, of leaf cells is responsible for elevated assimilate transport to the root. Increased sugar supply to the root, as observed in pgm mutants, forces substantial respiratory losses. Because root respiration accounts for 80% of total plant respiration under long‐day conditions, this gives rise to retarded biomass formation. In contrast, reduced vacuolar invertase activity leads to reduced net photosynthesis in the shoot and lowered root respiration, and affords an increased root/shoot ratio. The results demonstrate that roots have very limited capacity for carbon storage but exert rigid control of supply for their maintenance metabolism.