Efficient expression of human soluble guanylate cyclase in Escherichia coli and its signaling-related interaction with nitric oxide

Soluble guanylate cyclase (sGC), as a nitric oxide (NO) sensor, is a critical heme-containing enzyme in NO-signaling pathway of eukaryotes. Human sGC is a heterodimeric hemoprotein, composed of a α-subunit (690 AA) and a heme-binding β-subunit (619 AA). Upon NO binding, sGC catalyzes the conversion of guanosine 5′-triphosphate (GTP) to 3′,5′-cyclic guanosine monophosphate (cGMP). cGMP is a second messenger and initiates the nitric oxide signaling, triggering vasodilatation, smooth muscle relaxation, platelet aggregation, and neuronal transmission etc. The breakthrough of the bottle neck problem for sGC-mediated NO singling was made in this study. The recombinant human sGC β1 subunit (HsGCβ619) and its truncated N-terminal fragments (HsGCβ195 and HsGCβ384) were efficiently expressed in Escherichia coli and purified successfully in quantities. The three proteins in different forms (ferric, ferrous, NO-bound, CO-bound) were characterized by UV–vis and EPR spectroscopy. The homology structure model of the human sGC heme domain was constructed, and the mechanism for NO binding to sGC was proposed. The EPR spectra showed a characteristic of five-coordinated heme-nitrosyl species with triplet hyperfine splitting of NO. The interaction between NO and sGC was investigated and the schematic mechanism was proposed. This study provides new insights into the structure and NO-binding of human sGC. Furthermore, the efficient expression system of E. coli will be beneficial to the further studies on structure and activation mechanism of human sGC.     

中国野生鸭茅种质资源锈病抗性研究

鸭茅是一种著名的温带牧草,容易感染锈病。本研究从植株感病率、感病严重度、病情指数和反应指数4方面对国内37份鸭茅种质资源(野生资源35份)进行锈病的病情调查及抗病性研究。研究初步揭示了鸭茅锈病的发病规律,研究结果显示02-106、02-107、90-130、02-115等4份材料对鸭茅锈病具有较好抗性,可作为育种材料进一步深入研究。     

Expression analysis of miRNAs and highly-expressed small RNAs in two rice subspecies and their reciprocal hybrids

Heterosis,or hybrid vigor,is the phenomenon whereby progeny of two inbred lines exhibit superior agronomic performance compared with either parent.We analyzed the expression of miRNAs and highly expressed small RNAs(defined according to Solexa sequencing results)in two rice(Oryza sativa)subspecies(japonica cv.Nipponbare and indica cv.93-11)and their reciprocal hybrids using microarrays.We found that of all the 1141 small RNAs tested,140(12%,140 of 1141)and 157(13%,157 of 1141)were identified being significantly differentially expressed in two reciprocal hybrids,respectively.All possible modes of action,including additive,high- and low- parent,above high- and below low-parent modes were exhibited.Both F1 hybrids showed non-additive expression patterns,with downregulation predominating.Interestingly,15 miRNAs displayed stark opposite expression trends relative to midparent in reciprocal hybrids.Computational prediction of targets of differentially expressed miRNAs showed that they participated in multifaceted developmental pathways,and were not distinguishable from the targets of non-differentially expressed miRNAs.Together,our findings reveal that small RNAs play roles in heterosis and add a new layer in the understanding and exploitation of molecular mechanisms of heterosis.     

经脐单孔腹腔镜胆囊切除术的临床研究

目的:评价经脐单孔腹腔镜胆囊切除术在临床上应用的安全性及可行性.方法;回顾性分析2008年10月到2009年8月间我院收治的6例经脐单一切口腹腔镜胆囊切除手术患者的临床资料,并与同期15例传统腹腔镜胆囊切除术患者进行比较.结果:所有腔镜手术均成功,无术后并发症,恢复较好.经脐手术组的平均住院时间为7.33±0.82,与传统腔镜组的平均住院时间(6.73±0.96)无明显差异(P＞0.05);两者的平均住院费用也无差异(P＞0.05).结论:在慎重选择病例的基础上,经脐单孔腔镜胆囊切除术是一种安全、可行的手术.     

青少年双生子体型指征相关指标的遗传度研究(英文)

目的:用经典的双生子设计探讨遗传因素对青少年身高、体重、体质指数(BMI)等体型指征的影响程度。方法:从青岛市双生子库中选择自愿参加本研究的362对11-19岁双生子,测量身高、体重。同性别的双生子通过16个多态标记进行卵型鉴定,在此基础上,应用Mx软件构建结构方程模型分析计算遗传度。结果:对362对有效双生子数据进行分析,其中同卵双生子194对,异卵双生子168对,身高的最佳拟合模型ACE模型,体重和BMI的最佳拟合模型为AE模型。调整年龄性别后,身高的遗传度为66%,体重的遗传度为84%,体质指数的遗传度为75%。结论:在青少年体型指征的相关指标中,身高、体重、BMI受遗传因素影响都较大。     

Metabolism of organic acids, nitrogen and amino acids in chlorotic leaves of ‘Honeycrisp’ apple (Malus domestica Borkh) with excessive accumulation of carbohydrates

Metabolite profiles and activities of key enzymes in the metabolism of organic acids, nitrogen and amino acids were compared between chlorotic leaves and normal leaves of ‘Honeycrisp’ apple to understand how accumulation of non-structural carbohydrates affects the metabolism of organic acids, nitrogen and amino acids. Excessive accumulation of non-structural carbohydrates and much lower CO₂ assimilation were found in chlorotic leaves than in normal leaves, confirming feedback inhibition of photosynthesis in chlorotic leaves. Dark respiration and activities of several key enzymes in glycolysis and tricarboxylic acid (TCA) cycle, ATP-phosphofructokinase, pyruvate kinase, citrate synthase, aconitase and isocitrate dehydrogenase were significantly higher in chlorotic leaves than in normal leaves. However, concentrations of most organic acids including phosphoenolpyruvate (PEP), pyruvate, oxaloacetate, 2-oxoglutarate, malate and fumarate, and activities of key enzymes involved in the anapleurotic pathway including PEP carboxylase, NAD-malate dehydrogenase and NAD-malic enzyme were significantly lower in chlorotic leaves than in normal leaves. Concentrations of soluble proteins and most free amino acids were significantly lower in chlorotic leaves than in normal leaves. Activities of key enzymes in nitrogen assimilation and amino acid synthesis, including nitrate reductase, glutamine synthetase, ferredoxin and NADH-dependent glutamate synthase, and glutamate pyruvate transaminase were significantly lower in chlorotic leaves than in normal leaves. It was concluded that, in response to excessive accumulation of non-structural carbohydrates, glycolysis and TCA cycle were up-regulated to “consume” the excess carbon available, whereas the anapleurotic pathway, nitrogen assimilation and amino acid synthesis were down-regulated to reduce the overall rate of amino acid and protein synthesis.     

北江间(鱼骨)的繁殖生物学

间(鱼骨)(Hemibarbus medius Yue,1995)属鲤形目,鲤科,(鱼句)亚科,(鱼骨)属,为1995年报到的一个新种[1].在此之前,学者一直将其归为唇(鱼骨)[2-5].间(鱼骨)广泛分布于华南及西南部分地区,如珠江、南盘江、乌江和万泉河等水系[6].其肉质鲜嫩、营养价值高、产量较大,一直为产区的重要经济鱼类.     

连栽桉树人工林生产力和植物多样性及其相关性分析

对不同连栽代次的尾巨桉人工林进行长期定位观测,研究林分各组成部分生物量及林下植物多样性的变化,并采用趋势面分析方法分析生物量与植物多样性之间的关系.结果表明:(1)第1、2、3代7年生尾巨桉人工林的群落生物量基本相同,分别为102.22、105.11、102.66 t/hm2,其中乔木层林分生物量随着连栽代次呈增加趋势,第1代到第3代林分分别为81.48、88.36、92.28 t/hm2,平均净生产力分别为11.64、12.62、13.18 t/hm2·a,林分各器官生物量分配无明显差异,但在各生长年份,随着代数增加也呈增加趋势.(2)尾巨桉林下植物及枯落物生物量明显下降,第1、2、3代灌木层生物量分别为8.47、6.89、3.09 t/hm2,草木层生物量分别为1.93、1.21、0.35 t/hm2,枯落层生物量分别为10.34、8.64、6.94 t/hm2.(3)连栽尾巨桉人工林植物多样性指数、均匀度指数、丰富度指数随着连栽代次的增加而明显下降,即林下植物多样性随连栽代次的增加而减少,物种生态优势度增加.(4)采用趋势面分析方法结果表明,不同连栽代次尾巨桉林地植物多样性增加则林地根系生物量和林下植物生物量显著增加,而对林分总生物量及其它器官生物量影响不大.     

不明原因男性不育人群中睾丸特异性乳酸脱氢酶基因突变的发现及其意义

为了研究睾丸特异性乳酸脱氢酶,即乳酸脱氢酶C4(LDH-C4)基因突变在男性不育发病中的作用,利用LDH-C4特异性底物对100名不明原因男性不育症患者的精子LDH-C4进行活性显色,用变性高效液相色谱(DHPLC)技术对LDH-C4活性低下的患者进行LDHC基因PCR产物的突变筛查,对DHPLC峰形异常的PCR产物进行序列测定.筛选到一组精子LDH-C4活性明显下降的患者,其中1名患者的LDHC基因PCR产物在DHPLC中呈异常洗脱峰.对这一PCR产物进行序列测定,发现患者LDHC基因第5外显子的115位碱基发生了T→A的杂合改变(GenBank登录号GU479375),该突变使LDHC基因的178位密码子由原来的TTG(编码亮氨酸)变为TAG(终止密码子),形成截短的C亚基.T克隆-测序进一步证实了该无义突变的杂合状态.这是在人类LDHC基因上发现的第一个突变,提示LDHC基因突变可能是男性不育发病的原因之一.