Application of PCR-HRM in the study of relationship between leptin gene promoter polymorphisms and liver cirrhosis

AIM: To analyze the feasibility of PCR-high-resolution melting curve analysis (HRM) for detecting the site mutations of C2549 and G2548 in leptin gene promoter from the patients with liver cirrhosis, and to explore the relevance between mutant genotypes and physiological and biochemical indexes in liver cirrhosis patients. METHODS: Compared with the method of PCR-restriction fragment length polymorphism (RFLP), the present research used the method of PCR-HRM to analyze the site mutations of C2549 and G2548 in leptin gene promoter in control group (n=100) and liver cirrhosis group (n=100). The physiological and biochemical indexes of the patients were also detected and compared. RESULTS: Leptin gene promoter polymorphism was detected using PCR-HRM with effectiveness, high flux and accuracy. Preliminary results showed that the main mutation of the patients with liver cirrhosis was in C2549 site, but not found in G2548 site. Leptin, free leptin index (FLI), fasting insulin (FINS) and insulin resistance index estimated by homeostatic model assessment (HOMA-IR) in liver cirrhosis group were higher than those in control group. Insulin sensitivity index (ISI) and soluble leptin receptor (sOB-R) in liver cirrhosis group were lower than those in control group with significant difference except leptin level. Meanwhile, FLI showed positively correlated with FINS and HOMA-IR (r=0.45, r=0.53, P<0.05), and negatively with ISI (r=-0.34, P<0.05). In the patients with liver cirrhosis, C2549A heterozygous mutation was predominant. The indexes of HOMI-IR, leptin, sOB-R and FLI of C2549A homozygotes and heterozygotes were higher than those of the wildtypes, which showed significant difference except leptin and sOB-R levels (P<0.05). CONCLUSION: PCR-HRM can be more accurate for identifying leptin promoter polymorphism. The increase in the frequency of C2549A mutation may be closely related with liver cirrhosis. Existence of hyperinsulinemia and insulin resistance may be correlated with leptin level in the patients with liver cirrhosis.     

人心果转录组De novo组装及奇可胶途径相关基因的分析

为了探索人心果(Manilkara zapota L.)的转录组及奇可胶合成相关的基因,使用Illumina测序平台,对人心果果实、树皮和叶片分别进行转录组测序,使用Trinity等软件进行De novo组装和注释以及计算表达量,采用实时荧光定量PCR对转录组数据进行验证。经过组装得到162 455条unigene,总长度达139 792 553 nt,平均长度达861 nt,N50达1 544 nt。通过与NR、NT、Swiss-Prot、KEGG、COG和GO等数据库比对,共计89 628条unigene得到注释。以组装出来的unigene作为参考序列,在人心果转录组中共鉴别出57 362个SSR位点,果实、叶片和树皮表达的基因中分别检测到99 925、65 989和129109个SNP位点。在人心果转录组中共鉴别出105个与奇可胶合成相关的unigene,参与甲羟戊酸(MVA)途径的基因在果实和树皮中表达量较高,磷酸盐(MEP)途径中的基因则在叶片中的表达量较高,实时荧光定量PCR结果与转录组计算得到的表达量一致。初步推测MVA途径可能是奇可胶合成的主要途径。     

攀杂丝瓜1号新品种选育

攀杂丝瓜1号是以从重庆九龙坡丝瓜中选育的S8-6自交系为母本,以选自上海香丝瓜的自交系S4-3为父本,培育的早熟、丰产杂一代新品种,该品种植株生长势、分枝性强,坐果率高,果实长圆柱形,匀称,皮色深绿,表皮皱,瓜纵径26.5cm,横径5.5cm,单瓜质量285.8g,商品性好,味清甜,品质优,667m2产量2918.5kg。     

Relationship between airway inflammation and pulmonary arterial remodeling in rats with chronic bronchitis and emphysema

AIM:To study the effects of airway and pulmonary inflammation on pulmonary arterial remodeling in rats with chronic bronchitis (CB) and emphysema.METHODS:Twenty-four male Wistar rats were divided into three groups (n=8): Group A: four-weeks CB and emphysema;Group B: six-weeks CB and emphysema group;Group C: normal control.The rat model of CB and emphysema was established by intratracheal instillation of lipopolysaccharide (LPS) and daily exposure to cigarette smog.The arterial blood gas analysis,pulmonary hemodynamics changes and cell counts in bronchoalveolar lavage fluid (BALF) were measured.The pathomorphological changes of airway inflammation,alveoli destruction and pulmonary arterial remodeling were observed by HE straining and triple straining.RESULTS:(1) The characteristic pathological changes of CB and emphysema were observed in group A and B.Neutrophils were the main cells infiltrated into the walls of airway in group A.Lymphocytes and macrophages were the main cells in group B.(2) Right ventricular systolic pressure (RVSP),mean pulmonary arterial pressure (mPAP),the ratio of the weight of right ventricle/left ventricle and septum (RV/LV+S) in group A and B were significantly higher than those in group C (P<0.05).The amount of muscular artery (MA) in group A and B were significantly higher than that in group C (P<0.05).(3) In group A and B,the levels of MA,RVSP,mPAP and RV/LV+S was correlated positively with the average alveolar area,the total cell counts and differential cell counts of neutrophils,lymphocytes and macrophages in BALF,and the level of infiltration into the walls of airway,respectively (P<0.05).The positive correlation was observed with the percentage of neutrophils,lymphocytes and macrophages between group A and B (P<0.05).The amounts of MA were also correlated positively with RVSP,mPAP and RV/LV+S (P<0.05).CONCLUSIONS:(1) The pulmonary artery hypertension,the right ventricular hypertrophy and the pulmonary arterial remodeling appeared before hypoxia.These may be related with the degree of the pulmonary inflammation.(2) The characteristic of pulmonary arterial remodeling was small artery organization,and correlated positively with the changes of hemodynamics.     

噬菌体展示技术及其在抗血吸虫疫苗研究中的应用

噬菌体展示技术是新近发展起来的将外源肽或蛋白与特定噬菌体衣壳蛋白融合,并展示于噬菌体表面的一项新技术。示系统主要包括丝状噬菌体展示系统、入噬菌体展示系统、T4噬菌体展示系统及T7噬菌体展示系统等。该技术在抗血吸虫疫苗研究上的应用主要集中在模拟血吸虫抗原位点和直接作为疫苗载体方面,而在分析血吸虫保护性抗原位点、发现新的功能蛋白或抗原基因方面的应用研究还有待加强。本文就噬菌体展示拉术及其在抗血吸虫疫苗研制中的应用进行了简要概述,为抗血吸虫疫苗的研究提供。     

陇东旱塬冬小麦复种饲草轮作系统产量和水分利用特征

在陇东黄土高原大田条件下,连续2年研究了冬小麦-饲草复种轮作系统的干物质产量和水分利用,研究采用的处理有:休闲-冬小麦-箭筈豌豆-冬小麦(F-W-V-W),休闲-冬小麦-饲用油菜-冬小麦(F-W-R-W),饲用油菜-冬小麦-箭筈豌豆-冬小麦(R-W-V-W)和饲用油菜-冬小麦-饲用油菜-冬小麦(R-W-R-W)。结果表明:麦茬复种饲草可比麦茬休闲提高系统生产力27%,蛋白质产量增加1100 kg/hm², 同时对后茬冬小麦产量无影响。R-W-R-W模式下干物质产量、粗蛋白产量、干物质水分利用效率和蛋白质水分利用效率均最优,分别为30.68 t/hm²、3.25 t/hm²、7.80 kg/(mm·hm²)和0.92 kg/(mm·hm²)(P<0.05)。复种饲用油菜较箭筈豌豆产量高,有效利用了麦茬后降雨量,但箭筈豌豆粗蛋白含量较饲用油菜高22.4%,因此确定麦茬复种模式则需权衡干物质产量与蛋白质产量的需求。     

北川白山羊基因组与表型性状相关的多态性RAPD标记

北川白山羊是在特定生态条件下未经系统繁育而形成的优良地方山羊品种,在生长发育、繁殖和抗逆等性状方面具有突出优势。通过对122只北川白山羊个体基因组的扩增,结果从80条RAPD引物中选出12条多态引物,北川白山羊群体平均遗传相似率和遗传多样性指数分别为0.859 8±0.077 6和0.919 8,表明北川白山羊品种具有一定的遗传分化和较丰富的遗传多样性。在12条多态性引物中,SBS06同时对体质量（P=0.028）、体高（P=0.017）和体长（P=0.037）具有显著影响,SBS02对体质量（P=0.033）和体高（P=0.034）具有显著效应,所以推断影响体质量和体高性状的QTL基因座可能与RAPD标记SBS06和SBS02相连锁,可以应用于北川白山羊体质量和体高性状的标记辅助选择以及相应主效基因的进一步研究。     

桑树省力化全年条桑收获法的研究

全年条桑收获留干 0 4～ 0 .6m ,一留多年 ,且全年收获条桑。经过小区近 10年的试验及几年的农村示范表明 ,该收获法的收获工效比收获叶片提高 2倍以上 ,小区 9年平均产量比传统的冬根刈收获法增长 9 7个百分点 ,条桑饲养大蚕成绩明显比片叶饲养好。留干几年后根刈 ,桑树的再生长不受影响     

饲喂油菜籽对放牧牦牛乳生产性能和乳脂肪酸的影响

本试验主要目的是研究饲喂经高温高压处理的油菜籽对放牧牦牛乳生产性能和乳脂肪酸的影响.选择15头母牦牛,随机分为3组:对照组(N组,放牧);低饲喂量组(L组,放牧加饲喂油菜籽100 g/yak·d)和高饲喂量组(H组,放牧加饲喂油菜籽200 g/yak·d).结果表明:(1)与对照组相比,饲喂100 g/yak·d油菜籽对牦牛的乳生产性能和乳脂肪酸没有影响,饲喂200 g/yak·d油菜籽能显著提高乳产量以及各乳成分的产量(P<0.05);(2)棕榈酸(C16:0)、硬脂酸(C18:0)和油酸(c-9-C18:1)是放牧牦牛乳脂中含量较高的3种脂肪酸,饲喂油菜籽显著提高乳脂中硬脂酸(C18:0)的含量(P<0.05);(3)N、L和H组中共轭亚油酸(conjugated linoleic acid,CLA)含量分别是1.23%,1.15%和1.18%,饲喂油菜籽对各组CLA的含量和脱饱和酶指数无显著影响,牦牛乳中c-9,t-11-CLA的含量较高,平均为总CLA含量的98.60%.     

土壤Cu^2＋污染对植物生长影响的研究进展

工农业生产的飞速发展,导致土壤中的重金属越来越多。Cu^2＋是植物必需的金属营养元素之一,但过量会对植物产生一系列的毒害作用。因此,有关Cu^2＋胁迫对植物的毒害方面的研究成为人们关注的焦点之一。综述了土壤Cu^2＋污染对植物种子萌发、植株生长和生理效应3个方面的毒害作用。