基因编辑动物模型在人类疾病研究中的应用

近年来,随着以CRISPR/Cas9为代表的多种CRISPR系统的开发和不断改进,基因编辑技术逐渐完善,并广泛应用于人类疾病动物模型的制备。基因编辑动物模型为人类疾病的发病机理、病理过程以及预防和治疗等方面的研究提供了重要的素材。目前,用于人类疾病研究的基因编辑动物模型主要有小鼠、大鼠为代表的啮齿类动物模型和以猪为代表的大动物模型。其中啮齿类动物在机体各方面与人类差别较大,且寿命短,无法对人类疾病的研究和治疗提供有效评估和长期追踪;而猪在生理学、解剖学、营养学和遗传学等各方面与人类更接近,是器官移植和人类疾病研究领域重要的动物模型。文中主要介绍了基因编辑动物模型在神经退行性疾病、肥厚心肌病、癌症、免疫缺陷类疾病和代谢性疾病等5种人类疾病研究中的应用情况,以期为人类疾病研究及相关动物模型的制备提供参考。     

果胶甲酯酶的结构与功能研究进展

果胶甲酯酶(PME)是一种重要的果胶酶,其水解果胶中的甲酯基从而释放甲醇并降低果胶的甲酯化程度。目前在食品加工、茶饮料、造纸等生产工艺中有着广泛的应用前景。随着对PME的深入研究,已报道了几种不同来源的酶晶体结构,对这些已获得的晶体结构进行分析发现,PME属于右手平行β-螺旋结构,其催化残基为2个保守的天冬氨酸和1个谷氨酰胺残基,并且在催化过程中分别起到了一般酸碱、亲核试剂以及稳定中间体的作用。同时对其底物特异性进行分析,初步了解其底物与活性位点的识别机制。文中针对这几个相关方面进行了系统的综述。     

上海城市近自然森林的重建动态

近自然森林作为城市植被恢复的重要模式,已在我国多地开展了实践,并衍生了新的造林模式。为评估不同近自然森林建设模式的群落恢复进程,本研究以上海城市裸地上重建的近自然森林为对象,通过长期监测两处分别应用原"宫胁法"与新"异龄复层落叶—常绿混交林"种植模式的近自然森林重建过程,从物种组成、垂直结构、生活型组成和目标恢复物种4个方面解析恢复动态。结果表明:两种造林模式恢复的近自然森林,随恢复进程其物种组成逐渐趋同,在十多年内已形成了落叶—常绿垂直混交结构;"异龄复层落叶—常绿混交林"造林模式可更好地促进常绿阔叶树种的恢复,尤其是常绿建群种红楠与小叶青冈。本研究证实了近自然森林恢复技术可以缩短亚热森林群落向演替后期发展的时间,以及新造林模式的有效性,为近自然森林技术的应用与实施提供了科学依据。     

盐胁迫下菊芋根系脱落酸对钠离子转运和光系统Ⅱ的影响

通过根系施加脱落酸(ABA)合成抑制剂钨酸钠,研究盐胁迫(150 mmol·L^-1 NaCl)下菊芋根系ABA信号对根系Na⁺转运、叶片Na⁺积累和光系统Ⅱ(PSⅡ)的影响。结果表明:钨酸钠抑制盐胁迫下根系ABA合成,降低根系Na⁺外排,提高根系Na⁺向叶片的转运系数。盐胁迫增加叶片Na⁺含量,没有影响叶片膜脂过氧化、PSⅡ反应中心蛋白合成和PSⅡ最大光化学效率(F_v/F_m)。根系ABA合成受抑制,显著增加盐胁迫下叶片Na⁺积累,加剧叶片膜脂过氧化,损伤PSⅡ反应中心蛋白,显著降低F_v/F_m,诱发PSⅡ光抑制。总之,盐胁迫下菊芋根系ABA信号诱导根系Na⁺外排,抑制Na⁺向地上部转运,有利于减少叶片Na⁺积累,防御PSⅡ氧化损伤。     

UV-B辐射诱导芒果叶片抗氧化响应研究

为了解增强UV-B辐射诱导芒果叶片抗氧化响应的机制,以?台农1号?芒果(Mangifera indica?Tainong No. 1?)成年树为材料,以自然光为对照(CK),设置24和96 kJ/(m~2·d)两个增强UV-B辐射处理水平,观测叶片生理生化指标的动态变化。结果表明,24k J/(m~2·d)处理的芒果叶片MDA含量、相对电导率、净光合速率、抗氧化酶活性、多酚、Vc和芒果苷含量均与对照没有显著差异,而类黄酮和还原型GSH含量显著高于对照;而96 kJ/(m~2·d)处理的芒果叶片MDA含量、相对电导率、抗氧化酶活性及多酚、类黄酮、还原型GSH、芒果苷等还原型保护成分的含量均显著高于对照,而净光合速率和Vc含量均显著低于对照。因此,24 kJ/(m~2·d) UV-B辐射未引起?台农1号?芒果成年树损伤,可能是通过提高类黄酮和还原型GSH的含量来清除活性氧自由基;96 kJ/(m~2·d)处理则引起叶片活性氧损伤,但仍可能以两种机制减轻损伤,一是通过增强抗氧化酶活性和提高还原性成分含量来清除活性氧自由基,二是利用芒果苷、类黄酮和还原型GSH等成分吸收UV-B辐射。     

黄河三角洲河滩与潮滩芦苇对盐胁迫的生理生态响应

研究黄河三角洲两种生境芦苇对盐胁迫的生理生态响应差异,能为退化滨海湿地生态修复中芦苇植株来源的选择提供重要的理论支持。在盐胁迫(300 mmol/L NaCl)下,比较研究了黄河三角洲河滩芦苇(低盐生境)和潮滩芦苇(高盐生境)叶片中的Na~+含量、根部分生区Na~+流速、叶片的光合作用参数、H_2O_2的含量、抗氧化酶的活性、丙二醛和脯氨酸的含量。结果表明:盐胁迫显著提高了河滩芦苇叶片中Na~+含量,但对潮滩芦苇叶片Na~+影响不显著。进一步通过非损伤微测技术研究发现,盐胁迫后,潮滩芦苇比河滩芦苇的根部分生区的Na~+外排流速更高(潮滩芦苇:(1982.05±122.74) pmol cm~(-2) s~(-1)vs.(87.93±12.94) pmol cm~(-2) s~(-1),P0.01;河滩芦苇:(1574.16±458.90) pmol cm~(-2) s~(-1)vs.(-126.88±23.01) pmol cm~(-2) s~(-1),P0.01),能有效调节细胞内离子平衡。另外,盐处理7天后潮滩芦苇光合速率((16.36±1.09)μmol m~(-2) s~(-1)vs.(22.79±0.67)μmol m~(-2) s~(-1),P0.01)显著高于河滩芦苇((12.71±0.97)μmol m~(-2) s~(-1)vs.(23.81±0.55)μmol m~(-2) s~(-1),P0.01)。盐胁迫诱导了两种芦苇叶片中H_2O_2含量和丙二醛含量的显著升高,而脯氨酸及抗氧化酶的活性也随之显著升高,并且也显著升高了潮滩芦苇的谷胱甘肽还原酶(GR)活性((6.90±1.73) U/mg蛋白质vs.(3.54±0.54) U/mg蛋白质,P0.05)。综上,潮滩芦苇比河滩芦苇更适应盐胁迫,因其根系具有更高的排Na能力,且脯氨酸含量及抗氧化酶活性较高,提高了抗逆性。因此,在黄河三角洲退化滨海湿地生态修复中,可以优先选择潮滩芦苇作为植被修复的材料。     

农业主产区湖泊水质对湖滨带多尺度景观格局的空间响应

为分析农业主产区湖泊水环境质量对不同空间尺度上景观格局的响应关系,以洪湖为研究对象,以5种湖泊功能区为基础,利用RS和GIS软件生成7种空间尺度的湖滨带缓冲区,采用Fragstats 软件分析景观格局指数,结合冗余分析等数理统计方法与模型,研究景观格局对洪湖水质变化的空间尺度效应。结果表明: 1)景观格局在不同宽度缓冲区内对湖泊水质的影响具有空间尺度性,在200 m宽度其解释能力最大,达到86.1%,是影响水质的最有效空间尺度。2)景观配置变量(如最大斑块指数、斑块密度等)对水质的影响程度大于景观组成变量(如面积比例和均匀度指数等)。3)景观类型对水质的影响机制具有较大的差异性,农业用地受流域的地形地貌和耕作方式等因素的影响,在100～500 m小尺度的湖滨带缓冲区内对水质退化起主导作用;在远离水体的相对较大尺度(1000～5000 m),林地分布越密集、面积越大,对污染物进入水体的净化作用越明显;草地对水质的影响与林地一致;密集分布的城市用地对水质的影响与林地相反。研究结果可以从宏观尺度上为农业主产区湖泊流域水质管理和景观合理配置提供科学参考。     

Meta‐replication,sampling bias,and multi‐scale model selection: A case study on snow leopard (Panthera uncia) in western China

Replicated multiple scale species distribution models (SDMs) have become increasingly important to identify the correct variables determining species distribution and their influences on ecological responses. This study explores multi‐scale habitat relationships of the snow leopard (Panthera uncia) in two study areas on the Qinghai–Tibetan Plateau of western China. Our primary objectives were to evaluate the degree to which snow leopard habitat relationships, expressed by predictors, scales of response, and magnitude of effects, were consistent across study areas or locally landcape‐specific. We coupled univariate scale optimization and the maximum entropy algorithm to produce multivariate SDMs, inferring the relative suitability for the species by ensembling top performing models. We optimized the SDMs based on average omission rate across the top models and ensembles’ overlap with a simulated reference model. Comparison of SDMs in the two study areas highlighted landscape‐specific responses to limiting factors. These were dependent on the effects of the hydrological network, anthropogenic features, topographic complexity, and the heterogeneity of the landcover patch mosaic. Overall, even accounting for specific local differences, we found general landscape attributes associated with snow leopard ecological requirements, consisting of a positive association with uplands and ridges, aggregated low‐contrast landscapes, and large extents of grassy and herbaceous vegetation. As a means to evaluate the performance of two bias correction methods, we explored their effects on three datasets showing a range of bias intensities. The performance of corrections depends on the bias intensity; however, density kernels offered a reliable correction strategy under all circumstances. This study reveals the multi‐scale response of snow leopards to environmental attributes and confirms the role of meta‐replicated study designs for the identification of spatially varying limiting factors. Furthermore, this study makes important contributions to the ongoing discussion about the best approaches for sampling bias correction.     

丙酮丁醇梭菌(Clostridium acetobutylicum)原生质体融合育种研究

为了提高丙酮丁醇梭菌(Clostridium acetobutylicum)的丁醇耐受能力和培养基总糖产丁醇的转化率,通过原生质体融合的方法,研究了溶菌酶浓度及其作用时间、再生培养基种类、55℃条件下菌体致死时间、不同PEG分子量以及作用时间、Ca^2+和Mg^2+不同的添加量对丙酮丁醇梭菌原生质体制备、融合、再生的影响,得到了一套比较系统的丙酮丁醇梭菌的原生质体融合条件,同时通过气相色谱检测了融合菌的产溶剂能力并计算总糖转化率。结果显示,最终得到的215I菌株的总糖转化率比原始菌株提高了34.7%,产丁醇能力比原始菌株提高了32.2%,并且发现1株融合菌能产生新物质。原生质体融合方法在丙酸丁醇梭菌育种方面有广泛的应用潜力,通过融合得到的菌株为丁醇生产奠定了基础。     

Five candidate biomarkers associated with the diagnosis and prognosis of cervical cancer

Purpose: Cervical cancer (CC) is one of the most general gynecological malignancies and is associated with high morbidity and mortality. We aimed to select candidate genes related to the diagnosis and prognosis of CC.Methods: The mRNA expression profile datasets were downloaded. We also downloaded RNA-sequencing gene expression data and related clinical materials from TCGA, which included 307 CC samples and 3 normal samples. Differentially expressed genes (DEGs) were obtained by R software. GO function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs were performed in the DAVID dataset. Using machine learning, the optimal diagnostic mRNA biomarkers for CC were identified. We used qRT-PCR and Human Protein Atlas (HPA) database to exhibit the differences in gene and protein levels of candidate genes.Results: A total of 313 DEGs were screened from the microarray expression profile datasets. DNA methyltransferase 1 (DNMT1), Chromatin Assembly Factor 1, subunit B (CHAF1B), Chromatin Assembly Factor 1, subunit A (CHAF1A), MCM2, CDKN2A were identified as optimal diagnostic mRNA biomarkers for CC. Additionally, the GEPIA database showed that the DNMT1, CHAF1B, CHAF1A, MCM2 and CDKN2A were associated with the poor survival of CC patients. HPA database and qRT-PCR confirmed that these genes were highly expressed in CC tissues.Conclusion: The present study identified five DEmRNAs, including DNMT1, CHAF1B, CHAF1A, MCM2 and Kinetochore-related protein 1 (KNTC1), as potential diagnostic and prognostic biomarkers of CC.