茎尖嫁接脱除柑橘黄龙病病原及其检测方法比较

运用茎尖微芽嫁接技术脱除柑橘黄龙病病原,并采用PCR技术和直接荧光法对脱毒后的植株进行脱毒效率检测。结果表明,茎尖微芽嫁接成活率最高为75%;茎尖微芽嫁接技术是脱除黄龙病病原的一种有效方法,脱毒率达100%,脱毒后均检测不到病原存在;PCR技术检测病毒的检出率为100%,而直接荧光法的检出率仅为33.3%,说明PCR技术具有更高的灵敏度。     

Gene expression profile in immunologically injured liver cell of mice

To study the gene expression profiles between immunologically injured liver cell and normal liver cell of mice and to screen on a large scale the differentially expressed genes associated with the formation of liver injury, the experimental mice were randomly divided into the normal group for controlling and the immunologically liver-injured group induced by BCG and LPS. The liver mRNA of the two groups were extracted respectively and reversely-transcribed to cDNA with the incorporation of different fluorescence (Cy3, Cy5) labeled dUTP as the hybridization probes. The mixed probes were hybridized to the cDNA microarray chips. The fluorescent signal results were acquired by scanner ScanArray 4000 and analyzed with software GenePix Pro 3.0. Among the 14112 target genes, 293 genes were found to be significantly differentially expressed, in which 188 genes were up-regulated and 105 genes were down-regulated. Based on the analysis of biological functions of those differentially expressed genes, it was indicated that the occurrence and development of mouse liver damage induced by BCG and LPS were highly correlated with the processes of immune reactions, cell synthesis, metabolism, apoptosis and transportation in liver cell, which might be quite important for elucidating the regulatory network of gene expression associated with the liver damage, also important for finally discovering the pathogenic mechanisms of immunological liver damage.     

Wnt/β-catenin信号通路在博来霉素诱导的系统性硬化症小鼠模型血管重塑中的作用

系统性硬化症(systemic sclerosis,SSc)是一种慢性可累及全身多脏器的自身免疫性疾病,以广泛的血管病变及皮肤和内脏的纤维化为特征,但其机制迄今尚不明确。已有研究证实,Wnt通路参与了SSc纤维化,但其在血管病变中的病理作用尚未见报道。本研究拟采用博来霉素(bleomycin,BLM)诱导的SSc小鼠模型,探讨Wnt通路在SSc皮肤血管病变中的作用。将18只Balb/C小鼠随机平均分为3组,分别设为对照组(于小鼠背部皮下注射PBS 100 μL/d)、模型组(于小鼠背部皮下注射浓度为 1 mg/mL 博来霉素BLM 100 μL/d)和治疗组(于小鼠背部皮下注射 1 mg/mL BLM 100 μL/d,同时腹腔注射Wnt及β-catenin的抑制剂 iCRT3 5 mg/kg·d),于造模第28 d处死小鼠。小鼠皮肤取材后,通过HE染色及Masson染色观察到经BLM诱导的模型组小鼠背真皮、表皮厚度较对照组皮肤均明显增加(P<0.05),同时模型组的皮脂腺、毛囊等皮肤附属器明显减少,脂肪层厚度变薄并被纤维组织包绕,模型组皮肤胶原沉积较对照组增加;通过免疫组织化学染色在组织学层面鉴定α-SMA表达情况,发现模型组及治疗组α-SMA在皮肤组织中均高表达,α-SMA阳性表达在血管周围较对照组明显增加;通过ELISA方法检测出模型组小鼠血清中IL-6及IL-17表达量较对照组明显升高(P<0.05),治疗组小鼠血清中IL-6及IL-17的表达量较模型组明显下降(P<0.05);提取皮肤微血管片段,通过q-PCR检测到模型组及治疗组小鼠皮肤微血管中β-联蛋白的mRNA基因表达水平较正常组升高;通过Western印迹检测皮肤微血管Wnt5A、β-联蛋白、α-SMA、col1A1的蛋白质表达情况,发现纤维化相关蛋白质α-SMA及col1A1在模型组表达升高,较对照组有统计学差异(P<0.05),治疗组较模型组表达下降(P<0.05),Wnt通路相关蛋白质β-联蛋白及Wnt5A在模型组表达明显升高,较之对照组有统计学差异(P<0.05)。本研究提示,BLM能成功诱导小鼠系统性硬化症皮肤表型,Wnt通路的异常激活参与了BLM诱导的硬皮病小鼠皮肤微血管病变,特异性Wnt通路抑制剂iCRT3可能通过直接或间接的方式下调细胞因子IL-6及IL-17,从而降低BLM诱导的小鼠皮肤微血管中的α-SMA及col1A1蛋白质表达,改善小鼠皮肤微血管病变,干预BLM诱导的小鼠血管病变的进展。     

Successive chromosome walking by compatible ends ligation inverse PCR

Here we describe an advanced polymerase chain reaction (PCR) technique, the compatible ends ligation inverse PCR (CELI-PCR) for chromosome walking. In CELI-PCR, several restriction enzymes, which produce compatible cohesive ends, were used to digest target DNA simultaneously or sequentially to produce DNA fragments of suitable size. DNA fragments were then easily circularized and PCR amplification could be carried out efficiently. The previous limitations of inverse PCR were overcome, such as unavailable restriction sites, poor template DNA circularization, and low amplification efficiency. Therefore, successive chromosome walking was performed successfully. Our work, isolating a 11,395-bp fragment from Gossypium hirsutum, was presented as an example to describe how CELI-PCR was carried out.     

辽宁黄海海岸交错带的景观变化与设计

辽宁黄海海岸交错带的景观近几十来年变化很快,其中许多景观变化是有意识的人类活动或人工管理形成的景观设计可以看作是已经出现或即将出现的景观变化。由于这一区域海岸交错带有不同的海岸带类型,因而也就有了不同的景观设计方案。     

Tenascin-R distinct domains modulate migration of neural stem/progenitor cells in vitro

Extracellular matrix (ECM) molecules constitute a "niche" that modulates the migration, proliferation, and differentiation of neural stem/progenitor cells (NSPCs). The glycoprotein Tenascin-R (TN-R) is an ECM molecule, comprising multiple domains. Either the whole TN-R molecule or its distinct domains has been demonstrated to play a very important role in the developing central nervous system. However, little is known about the effect of the TN-R domain on NSPCs, especially NSPC migration. In the present study, we first show that both TN-R domains epidermal growth factor-like repeat (EGFL) and fibronectin type III (FN)6-8 can inhibit the NSPCs migration from neurospheres in vitro. Furthermore, both the EGFL and FN6-8 domains affect the distribution of neurons generated from neurospheres, indicating that EGFL and FN6-8 domains inhibit the motility of neurons generated from neurospheres. These results suggest that TN-R has an inhibitory effect on NSPCs migration.     

高油玉米优良自交系再生体系的建立

以供试的5个高油玉米优良自交系为材料,建立了一个高效的高油玉米幼胚再生体系.研究表明,高油玉米幼胚组织培养的最适幼胚长轴长度在0.5 mm～2.0 mm左右;MB培养基是最适的胚性愈伤组织诱导培养基;各材料胚性愈伤组织诱导率差异较大,以4K261和4K296的胚性愈伤诱导率较高;不同材料最适的继代培养条件存在差异,但基因型仍然是决定各自交系胚性愈伤组织的继代能力的主导因素,其中以4K261最佳.5个自交系均能分化出幼苗,但分化率差异较大,以4K059分化率最高,达82.0 %;其次是4K261和4K296,分别为63.2 %和59.0 %;4K060和4K061表现最差.所以4K059、4K261和4K296均可作为遗传转化的受体材料.该体系的建立为高油玉米的遗传转化奠定了基础.     

单穗升麻的柱头和雌配子体发育及胚胎发生

单穗升麻的雌蕊群由1～5枚离生心皮组成。子房单室,4～9枚倒生胚珠,双珠被,厚珠心;大孢子四分体呈线形或T形排列,合点端一个具功能。胚囊发育为蓼型。成熟胚囊中3个单核反足细胞,胞质浓密,在球形胚时期尚能观察到其退化痕迹。极核融合早,次生核位于合点端。胚胎发生为柳叶菜型;核型胚乳。雄蕊凋谢后1～2天花柱顶端腹缝线周围出现乳突细胞,雄蕊凋谢后3～5天延长成柱头毛。讨论了单穗升麻与南川升麻柱头可授期与花粉生活力的差异对传粉效果和结籽率的显著影响。     

中华绒螯蟹白化症、正常肝胰腺组织结构及脂肪酸组成的比较研究

采用气相色谱、组织学及电镜技术分别对中华绒螯蟹雄蟹白化症肝胰腺与正常肝胰腺脂类和脂肪酸组成、显微及超微结构进行了比较研究,结果表明：正常肝胰腺的总脂含量（31.54%肝胰腺湿重）显著高于白化症肝胰腺（19.07%肝胰腺湿重）,两种蟹肝胰腺的脂类组成没有显著差异,但脂肪酸组成存在一定的差异,白化症肝胰腺中C18∶1的含量显著高于正常肝胰腺,中性脂和极性脂HUFA正常蟹肝胰腺含量都高于白化症蟹,但没有显著差异。组织学结果和超微结构观察表明：白化症肝胰腺肝小管的上皮细胞壁结构紊乱,微绒毛及微绒毛黏膜大部分脱落,细胞内仅见有零星的线粒体、脂肪滴分布,R细胞呈现空泡化。