桔小实蝇线粒体基因组全序列及其分析

桔小实蝇Bactrocera dorsalis线粒体基因组全序列对研究实蝇分子系统进化具有重要意义。本研究通过DNA测序和克隆技术,对桔小实蝇mtDNA全序列进行了测定和分析。结果表明：桔小实蝇线粒体基因组全长15 915 bp（GenBank序列号: DQ845759）。基因组碱基组成为39.3%A,16.2%C,10.2%G,34.3%T,由13个蛋白编码基因、22个tRNA基因、2个rRNA基因以及一个非编码的控制区域（A+T-rich区）组成。7个蛋白编码基因和13个tRNA基因从J链编码,其余6个蛋白编码基因和9个tRNA基因从N链编码。位于J链上的蛋白编码基因具有近似的A、T含量,而位于N链上的蛋白编码基因的A的含量明显高于T的含量。以mtDNA COⅠ基因为例,比较了桔小实蝇与其他14种实蝇的亲缘关系,结果显示其与同亚属（果实蝇亚属Bactrocera）内的其他近缘种相互间的同源性很高。     

Acetone-soluble cellulose acetates prepared by one-step homogeneous acetylation of cornhusk cellulose in an ionic liquid 1-allyl-3-methylimidazolium chloride (AmimCl)

Cellulose samples extracted from cornhusk have been successfully acetylated in an ionic liquid 1-allyl-3-methylimidazolium chloride (AmimCl). Without using any catalyst, cornhusk cellulose acetates (CCAs) with the degree of substitution (DS) in a range from 2.16 to 2.63 were prepared in one-step. Under the homogeneous state, the DS value of CCAs was easily controlled by the acetylation time. The obtained CCAs were characterized by means of FT-IR, ¹³C NMR, DSC, TGA, and a mechanical test. The NMR results showed that the distribution of the acetyl moiety among the three OH groups of the anhydroglucose unit shows a preference at the C₆ position. The CCAs exhibited good solubility in some organic solvents, such as acetone and DMSO. The cast CCA films from their acetone solutions had good mechanical properties. At the end of each acetylation of cornhusk cellulose, the ionic liquid AmimCl could be effectively recovered. Therefore, this study presents a promising approach and “green process” to make use of crop by-products.     

地方稻资源D43的开花期耐热特性研究

水稻在抽穗开花期对高温胁迫非常敏感,通过挖掘耐热资源,培育耐热水稻品种是应对高温热害最有效的方式。前期研究发现,地方稻资源D43在花期连续高温条件下能保持较高的结实率。本研究在大田和人工气候室不同高温处理下,分析了D43的开花时间与耐热性之间的相互关系。结果表明,高温能够使水稻的开花时间提前,D43表现出稳定的早花时特性,高温胁迫下的开花时间集中在8∶30~10∶00;在开花时间段恒定高温胁迫下,D43的结实率较低;但在大田高温和人工气候室模拟高温胁迫下,D43的开花时间避开了日高温段,从而表现出较高的结实率;花器官形态性状包括花药开裂率、柱头上的花粉附着数、花粉萌发数与结实率之间呈显著正相关,因此可用于评价水稻的花期耐热性。     

3种滨藜属牧草苗期叶片解剖结构及生理特性对干旱的响应

通过盆栽控水试验,设置5个干旱胁迫水平,分别为最大田间持水量的80%(CK)、60%(轻度胁迫)、45%(中度胁迫)、30%(重度胁迫)、20%(极重度胁迫),并同步设计充足灌水后自然干旱实验,测定干旱胁迫对3种滨藜属牧草叶片形态解剖结构、叶片相对含水量、叶绿素含量、净光合速率、可溶性糖含量、质膜相对透性和丙二醛含量等生理生化指标的影响,以明确3种滨藜属牧草的抗旱特性,并探索其抗旱机理。结果表明:(1)3种滨藜属牧草均具有适应旱生环境的典型叶片结构特征,即在干旱胁迫条件下叶片组织结构形态表现为叶片栅栏组织逐渐变薄,而海绵组织在胁迫早期先变薄后增厚的现象,叶肉组织结构紧密度也出现了先降低后增高的规律。(2)随着干旱胁迫的加剧,叶片的可溶性糖含量增加,而其相对含水量减少,3种滨藜属牧草在土壤含水量很低的情况下叶片仍能保持高于52%的相对含水量。(3)在干旱胁迫下,3种滨藜属牧草叶片的叶绿素含量、净光合速率、气孔导度和蒸腾速率都发生显著变化,细胞膜受到的伤害程度有明显差异。(4)3种滨藜属牧草抗旱能力均较强,在干旱胁迫下其抗旱性综合表现为灰白滨藜变种1灰白滨藜变种2四翅滨藜。     

人胎盘滋养层细胞培养与体外hCG释放的研究

本研究的目的是了解细胞滋养层细胞和合胞体滋养层细胞体外分化和生物学特性。方法:采用酶消化和Percoll密度梯度离心法,对人足月胎盘细胞滋养层细胞进行分离、纯化和体外培养。采用放射免疫法(RIA)检测细胞培养上清液hCG含量的变化。结果:经分离和纯化的细胞滋养层细胞在体外培养中生长良好,通过细胞分裂和融合形成合胞体滋养层细胞,随着合胞体滋养层细胞的生长,细胞培养上清液中hCG含量显著升高。我们认为从胎盘中分离和纯化的细胞滋养层细胞在体外培养中可分化和融合形成合胞体滋养层细胞,体外hCG含量的增加与合胞体滋养层细胞生长有关。     

New mechanism of nerve injury in Alzheimer’s disease: β‐amyloid‐induced neuronal pyroptosis

The present study was designed to investigate the role of β‐amyloid (Aβ_1‐42) in inducing neuronal pyroptosis and its mechanism. Mice cortical neurons (MCNs) were used in this study, LPS + Nigericin was used to induce pyroptosis in MCNs (positive control group), and Aβ_1‐42 was used to interfere with MCNs. In addition, propidium iodide (PI) staining was used to examine cell permeability, lactate dehydrogenase (LDH) release assay was employed to detect cytotoxicity, immunofluorescence (IF) staining was used to investigate the expression level of the key protein GSDMD, Western blot was performed to detect the expression levels of key proteins, and enzyme‐linked immunosorbent assay (ELISA) was utilized to determine the expression levels of inflammatory factors in culture medium, including IL‐1β, IL‐18 and TNF‐α. Small interfering RNA (siRNA) was used to silence the mRNA expression of caspase‐1 and GSDMD, and Aβ_1‐42 was used to induce pyroptosis, followed by investigation of the role of caspase‐1‐mediated GSDMD cleavage in pyroptosis. In addition, necrosulfonamide (NSA), an inhibitor of GSDMD oligomerization, was used for pre‐treatment, and Aβ_1‐42 was subsequently used to observe the pyroptosis in MCNs. Finally, AAV9‐siRNA‐caspase‐1 was injected into the tail vein of APP/PS1 double transgenic mice (Alzheimer's disease mice) for caspase‐1 mRNA inhibition, followed by observation of behavioural changes in mice and measurement of the expression of inflammatory factors and pyroptosis‐related protein. As results, Aβ_1‐42 could induce pyroptosis in MCNs, increase cell permeability and enhance LDH release, which were similar to the LPS + Nigericin‐induced pyroptosis. Meanwhile, the expression levels of cellular GSDMD and p30‐GSDMD were up‐regulated, the levels of NLRP3 inflammasome and GSDMD‐cleaved protein caspase‐1 were up‐regulated, and the levels of inflammatory factors in the medium were also up‐regulated. siRNA intervention in caspase‐1 or GSDMD inhibited Aβ_1‐42‐induced pyroptosis, and NSA pre‐treatment also caused the similar inhibitory effects. The behavioural ability of Alzheimer's disease (AD) mice was relieved after the injection of AAV9‐siRNA‐caspase‐1, and the expression of pyroptosis‐related protein in the cortex and hippocampus was down‐regulated. In conclusion, Aβ_1‐42 could induce pyroptosis by GSDMD protein, and NLRP3‐caspase‐1 signalling was an important signal to mediate GSDMD cleavage, which plays an important role in Aβ_1‐42‐induced pyroptosis in neurons. Therefore, GSDMD is expected to be a novel therapeutic target for AD.     

Myokine IL-15 regulates the crosstalk of co-cultured porcine skeletal muscle satellite cells and preadipocytes

The present study was carried out to preliminarily reveal the underlying mechanisms of the co-culture system between porcine muscle satellite cells (SCs) and stromal-vascular cells (SVs). The two cell types were co-cultured to assess both proliferation and differentiation. Desmin and Pref-1 immunofluorescence staining technique were taken to identify the two types of isolated cells. The expression of specific marker genes Myogenin was up-regulated in SCs (P < 0.05) and the differentiation of SCs could be promoted when co-cultured with preadipocytes compared with the single-cultured control, while expression of c/EBPβ in SVs was down-regulated (P < 0.05) and the differentiation of preadipocytes could be inhibited. Furthermore, secretion of myokine IL-15 was markedly increased, as well as its gene and protein expression levels in co-culture supernatants. However, the secretion of adipokine leptin was significantly decreased. These findings demonstrate that myokines like IL-15 could facilitate the SCs’ differentiation while inhibit the SVs differentiation, and act as an important regulator of co-culture between muscle cells and adipocytes.     

脉冲电场对人红细胞膜结构影响的冷冻断裂研究

对外加脉冲电场处理的人红血球冷冻断裂和蚀刻的复型观察中发现在强电场(3KV/cm)作用下,细胞周围有颗粒状和纤维状结构。结合SDS电泳分析证明了它们是由于在电场作用下,红血球膜的带3蛋白和膜骨架蛋白(血影蛋白)脱出的结果。在强电场作用下,由于膜蛋白和膜骨架蛋白的脱出造成了对细胞膜的损伤,使细胞膜稳定性降低,细胞易变形和形成伪足。由于膜蛋白的脱出,多余的自由脂质进入细胞质内而形成泡状结构。外电场改变了蛋白-蛋白以及蛋白-脂分子间的作用可能是电穿孔的主要机理。本文还对当前公认的冷冻断裂中所观察到的膜中间颗粒的来源提出了疑问,并提出了它们还可能与冰晶有关。而冰晶的形成又与膜的亲水与疏水性有关。     

碱性成纤维细胞生长因子在苜蓿中的表达

为了降低bFGF(basic fibroblast growth factor)的生产成本,结合植物生物反应器的优点,就bFGF在转基因苜蓿中的表达进行了探索.将bFGF插入植物表达载体pBⅡ21中,获得了含有bFGF基因的植物表达pBIcbFGF,再将pBIcbFGF利用冻融法转到农杆菌中.利用农杆菌介导法将基因转化保定苜蓿,转基因苜蓿在TM-1培养基+20 mg/L卡那霉素(Kan)+200 mg/L特美汀(Tim)中诱导分化,在生根培养基中生根,获得再生植株.再生植株通过PCR检测、RT-PCR及Western blot证实外源基因已经在苜蓿中成功表达.获得含有目的蛋白的阳性植株.为苜蓿作为植物生物反应器生产bFGF奠定了理论及技术基础.