猪脂肪组织表达序列标签(ESTs)大规模测序及分析

利用大规模DNA序列测定的方法,对猪脂肪组织进行了表达序列标签(Expressed Sequence Tag,EST)序列测定,获得高质量EST共7790个,并对此进行了初步分析。使用STACK-PACK软件进行聚类分析,得到4354个基因聚类,包括3609个单拷贝基因和745个多拷贝基因;将候选基因序列用BlastN与nr库进行比较(e=1e-10),从4354个候选基因中得到2712个已知基因,其中单基因为1987个,多拷贝基因为725(3694克隆)个;未知功能基因和新EST有2109个克隆。根据BlastN结果,利用基因组文库添加序号(GenBank Accession No．)为索引,构建了猪脂肪组织已知功能基因表达谱。从基因表达谱可以看出,在猪脂肪组织中参与代谢的基因所占比例最高,在某些方面也显示了脂肪组织旺盛的代谢活性。同时发现在猪脂肪组织中主组织相容性抗原(Major Histocompatibility Complex,MHC)或与MHC相关的基因表达丰度很高。其中单拷贝基因181个,多拷贝基因44个,共计257个克隆,占细胞机体防御(cell and organism defense)总数的44．9％。占总已知基因数的5.4％。提取出全部与MHC相关的EST序列(257个克隆),发现所有EST的部分序列(长约200个碱基),几乎分布在每一个已知猪BAC的所有编码序列上。据此推测,构成MHC的这些EST序列中,有一段长约200个碱基(200bp)的碱基序列高度保守,MHC基因中每一段编码序列都包含有这一段序列。这些MHC序列虽然在不同的BAC上其蛋白的域不同,但均为高度保守区域,并且都与免疫功能密切相关。猪脂肪中如此大量表达的MHC部分保守序列,由于与免疫功能高度相关,在MHC基因的传递过程中,可以反复复制,并能够稳定遗传。     

石油焦基高比表面积活性炭对废水中CODCr的吸附能力

探讨了高比表面积活性炭(HSAAC)吸附水中COD_Cr时,活性炭用量、pH值和吸附时间等因素对COD_Cr吸附量和去除率的影响。实验结果表明,HSAAC用量越大,去除COD_Cr效果越好。当HSAAC用量为2.0g·L^-1,pH=3时,去除率达到78%以上;在酸性条件下HSAAC对COD_Cr的去除效果较好;HSAAC对废水中COD_Cr的吸附发生在前30min;COD_Cr浓度低于60mg·L^-1时,处理后COD_Cr的残余质量浓度低于地表水环境质量Ⅰ类标准(15mg·L^-1)。用碱再生HSAAC,一次再生率达94.22%,二次再生率达到了86.90%。说明高比表面积活性炭在适宜条件下对COD_Cr具有较好的吸附性能和良好的再生效果。     

雉科8种中央尾羽分类特征探究

观察348个标本的中央尾羽,测量其性状并统计,结果表明:1)雉科8种中央尾羽色斑格局不相同:2)中央尾羽长、内翈宽、外翈宽都存在显著差异(P<0.05),以白冠长尾雉中央尾羽(118,3±18,89)cm最长,内翈(2.44±0.21)cm最宽,以红腹锦鸡外翈(2.80±0.82)cm最宽,而灰胸竹鸡的中央尾羽(9.51±0.78)cm最短,内翈(1.02±0.13)cm和外嘲(0.99±0.10)cm最窄.以上有助于分辨雉科此8种鸟类.     

猴免疫缺陷病毒特异性免疫复合物沉着与猴AIDS多系统病变关系的探讨

目的观察猴免疫缺陷病毒(SIV)感染后,病毒特异性免疫复合物(IC)在不同时间、不同组织的沉着情况,初步研究其与AIDS多系统病变的联系。方法对8只不同时间感染SIV的恒河猴及1只未感染SIV的猴进行尸检以获取多系统组织标本,进行连续切片和IgG、C3、SIV p27免疫荧光染色,并对结果进行比较分析。结果IgG、C3、p27在多个猴、多种组织的相同位置出现相同模式的荧光表达,证明存在IC的沉着;其中脑血管周(8/8),心肌间微血管(6/8)、和淋巴结副皮质(6/8)及生发中心(5/8)是阳性率最高的部位,肾小球及肾间质、肠黏膜固有层也有较多IC的沉着,且在感染中、晚期IC出现的比例更高。结论SIV感染后出现广泛的SIV-IC沉积,且随病情的进展而加重;IC可能是SIV导致AIDS多系统病变的主要形式。针对此过程进行研究,可帮助了解AIDS并发多系统器官病变的机制及研究新的治疗方法。     

Fine mapping and candidate gene analysis of dense and erect panicle 3, DEP3, which confers high grain yield in rice (Oryza sativa L.)

Architecture of the rice inflorescence, which is determined mainly by the morphology, number and length of primary and secondary inflorescence branches, is an important agronomical trait. In the current study, we characterized a novel dense and erect panicle (EP) mutant, dep3, derived from the Oryza sativa ssp. japonica cultivar Hwacheong treated with N-methyl-N-nitrosourea. The panicle of the dep3 mutant remained erect from flowering to full maturation, whereas the panicle of the wild type plant began to droop after flowering. The dep3 mutation also regulated other panicle characteristics, including panicle length, grain shape and grain number per panicle. Anatomical observations revealed that the dep3 mutant had more small vascular bundles and a thicker culm than wild type plants, explaining the EP phenotype. Genetic analysis indicated that the phenotype with the dense and EP was controlled by a single recessive gene, termed dep3. The DEP3 gene was identified as the candidate via a map-based cloning approach and was predicted to encode a patatin-like phospholipase A2 (PLA2) superfamily domain-containing protein. The mutant allele gene carried a 408?bp genomic deletion within LOC_Os06g46350, which included the last 47?bp coding region of the third exon and the first 361?bp of the 3??-untranslated region. Taken together, our results indicated that the patatin-like PLA2 might play a significant role in the formation of vascular bundles, and that the dep3 mutant may provide another EP resource for rice breeding programs.     

Role of myosin light chain kinase in regulation of basal blood pressure and maintenance of salt-induced hypertension

Vascular tone, an important determinant of systemic vascular resistance and thus blood pressure, is affected by vascular smooth muscle (VSM) contraction. Key signaling pathways for VSM contraction converge on phosphorylation of the regulatory light chain (RLC) of smooth muscle myosin. This phosphorylation is mediated by Ca(2+)/calmodulin-dependent myosin light chain kinase (MLCK) but Ca(2+)-independent kinases may also contribute, particularly in sustained contractions. Signaling through MLCK has been indirectly implicated in maintenance of basal blood pressure, whereas signaling through RhoA has been implicated in salt-induced hypertension. In this report, we analyzed mice with smooth muscle-specific knockout of MLCK. Mesenteric artery segments isolated from smooth muscle-specific MLCK knockout mice (MLCK(SMKO)) had a significantly reduced contractile response to KCl and vasoconstrictors. The kinase knockout also markedly reduced RLC phosphorylation and developed force. We suggest that MLCK and its phosphorylation of RLC are required for tonic VSM contraction. MLCK(SMKO) mice exhibit significantly lower basal blood pressure and weaker responses to vasopressors. The elevated blood pressure in salt-induced hypertension is reduced below normotensive levels after MLCK attenuation. These results suggest that MLCK is necessary for both physiological and pathological blood pressure. MLCK(SMKO) mice may be a useful model of vascular failure and hypotension.     

MicroRNAs and their role in viral infection

Recently, a class of about 22 nucleotides (nt) small RNA has been discovered in many eukaryotes, termed microRNAs (miRNAs), which have a variety of functions. Many recent findings have demonstrated that viruses can also encode their own miRNAs. Meanwhile, other findings reveal a relationship between host miRNA and viral infection. These findings suggest a tight relationship between host and viral infection via miRNA pathway. This article introduces the miRNAs encoded by viruses and reviews the advances of the interaction of the mammalian host miRNAs and viral infection.     

半抗原特异性抗体的筛选及亲和力成熟

噬菌体抗体库技术的出现开创了一条便捷的基因工程抗体生产路线,为小分子半抗原抗体的制备提供了一条新途径,具有极大的应用潜力,但得到的抗体片段的亲和力普遍较全分子差,需要优化筛选策略及抗体的体外亲和力成熟来解决这一问题。     

CRL4在增殖的肝细胞系中上调乙型肝炎病毒抗原的分泌表达

乙型肝炎病毒（hepatitis B virus,HBV）编码的X蛋白（hepatitis B virus X protein,HBx）对HBV感染的起始和维持至关重要。HBx可能作为病毒来源的接头分子,介导Cullin-RING E3泛素连接酶4（cullin-RING ubiquitin E3 ligase 4,CRL4）复合物对染色体外DNA限制因子SMC5/6的降解。最近研究发现CRL4接头分子DNA损伤结合蛋白1（DNA damage-binding protein 1,DDB1）可不依赖与HBx的相互作用而直接上调病毒的表达和复制。本研究基于HBx基因删除（X-null）的HBV重组cccDNA（recombinant covalently closed circular DNA,rcccDNA）模型系统,在多种体外培养肝细胞系中证实上述发现。有意思的是,CRL4刺激rcccDNA^X-null转染细胞抗原分泌表达的效应能被血清饥饿实验抵消。应用尾静脉高压注射小鼠模型,同样发现CRL4并不上调rcccDNA^X-null在非增殖小鼠肝脏细胞中的表达。以上结果提示,细胞增殖特征与CRL4不依赖HBx上调病毒抗原表达的效应密切相关,有助于HBx生物学意义的准确分析和理解。     

阿替普酶联合依达拉奉治疗急性缺血性卒中的疗效及神经功能缺损与时间窗的关系研究

目的:探讨阿替普酶联合依达拉奉治疗急性缺血性卒中的疗效及神经功能缺损与时间窗的关系。方法:选取大连医科大学附属大连市中心医院于2016年3月~2018年10月间收治的急性缺血性卒中患者117例,根据随机数字表法将患者分为对照组(n=58,阿替普酶治疗)和研究组(n=59,阿替普酶联合依达拉奉治疗),比较两组患者临床疗效、神经功能缺损情况、基质金属蛋白酶-9(MMP-9)、白介素-6(IL-6)水平、头颅CT梗死面积,观察两组治疗期间不良反应发生情况。结果:研究组的总有效率为84.75%(50/59),高于对照组的63.79%(37/58)(P<0.05)。两组患者治疗2周后MMP-9、IL-6、美国国立卫生研究院卒中量表(NIHSS)评分均较治疗前降低,且研究组低于对照组(P<0.05)。研究组治疗24h、48h、72h的头颅CT梗死面积小于对照组(P<0.05)。治疗后研究组发病72h内、发病48h内患者NIHSS评分、头颅CT梗死面积高于发病24h内,且发病72h内高于发病48h内(P<0.05)。两组患者不良反应发生率比较无差异(P>0.05)。结论:阿替普酶联合依达拉奉治疗急性缺血性卒中,疗效确切,可有效改善患者过氧化损伤,降低细胞因子水平,且越早的时间窗内接受治疗的患者,其神经功能缺损、脑梗死面积改善效果越好。