High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

五种鬼伞过氧化物酶和酯酶的同工酶研究

应用垂直板聚丙烯酰胺凝胶电泳对五种野生鬼伞 (Coprinus)真菌进行了过氧化物酶 (POD)和酯酶(EST)的同工酶分析 ,结果表明 :五种鬼伞的POD和EST同工酶酶谱比较稳定清晰 ,且分别有一条共同的酶带 ,可能是鬼伞属的POD和EST同工酶特征酶带 ;POD和EST同工酶酶谱均表明 ,家园鬼伞 (C .domesticus)和瓦鳞鬼伞 (C .clavatus)间有较近的亲缘关系 ;不同种鬼伞的POD和EST同工酶之间既有共同的特征 ,又各自有本物种的特有特征 ,POD和EST同工酶酶谱可以作为鬼伞属种类鉴定、亲缘关系比较的重要依据。     

Roles of the Kv1.2, Kv1.5, Kv2.1 potassium channels in hypoxia pulmonary vasoconstriction

AIM: To determine the role of Kv1.2, Kv1.5, Kv2.1 in the hypoxia pulmonary vasoconstriction (HPV). METHODS: Male Wistar rats were divided into two groups: normoxic group and hypoxic group. The single smooth muscle cell was obtained from pulmonary artery of Wistar rats with acute enzymatic digestion method. The conventional whole-cell patch clamp technique was used to record the resting membrane potential (Em) and the potassium currents of voltage-gated potassium channel (IKv) in rat pulmonary arterial smooth muscle cells (PASMC). Intracellular application of Kv1.2/Kv1.5/Kv2.1 antibodies (1∶125) was conducted through the whole-cell patch clamp system. RESULTS: ① Em of PASMC was depolarized after 24 h hypoxia compared with that of control cells . IKv of PASMC was decreased after 24 h hypoxia, . ② The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies depolarized Em and inhibited IKv in PASMC from normoxic rat, whereas the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on them. ③ The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies and the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on IKv and Em from rats hypoxic for 24 h. CONCLUSION: Kv1.2, Kv1.5, Kv2.1 might be oxygen sensitive potassium channels which mediated HPV.     

In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time‐period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital^® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post‐thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital^® (SV) technology, the first commercialized production line of SpermVital^® (C) and by conventional procedure applying Biladyl^® extender (B). Post‐thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital^® technology is improved and is more efficient in conserving post‐thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.     

菱传粉生物学研究

以6 个菱品种为试材，对花器形态特征、群体花期及单花开花动态、开花时间、柱头可授性、花粉活力、杂交指数 及传粉形式等进行观察测定。结果表明：菱花单生，两性，单花花期15 h，群体花期在5～10 月，盛花期在7～8 月；不同 品种在一天中的开花高峰期存在差异；柱头可授期主要在萼片开裂后2 h 内，结实率在50% 以上；花粉在自然授粉条件下活 力可保持7.5 h；以自花授粉为主，同时存在一定程度的无融合生殖和少量异花授粉。     

一种高质量天门冬基因组DNA提取方法

针对天门冬[Asparagus cochinchinensis(Lour.)Merr.]成熟叶片富含多糖、多酚、蛋白质及其他次生代谢物质的特点,采用改良CTAB法对其基因组总DNA进行提取,并对DNA进行质量检测。结果表明,改良CTAB法可从天门冬成熟叶片中获得纯度高、完整性好的总DNA,其OD260 nm/OD280 nm为1.86,OD260 nm/OD230 nm为2.36,浓度为390μg/mL,产率达11.7μg/g,多糖、多酚和RNA杂质被去除干净,无降解现象,说明该方法提取的天门冬基因组总DNA质量较高。     

渗灌次数对西北荒漠灌区紫花苜蓿栽培草地土壤化学性质的影响

渗灌是节水农业中最有效的措施之一,有利于作物产量和水分利用率的提高。本研究以甘肃省金昌市杨柳青公司紫花苜蓿‘艾迪娜’(M.sativa L.‘Aidina’)试验地渗灌区土壤为研究对象,通过田间试验与室内分析相结合的方法,研究不同渗灌次数(0次、1次、2次、3次)对苜蓿出苗期不同土层深度及管距位置土壤化学性质的影响。结果表明:在0~10cm和10~20cm土层深度,pH最高值均出现在管上,较CK升高了8.21%和8.68%,最低值都出现在管中,较CK降低了2.70%和3.37%;2水和3水均能显著提高不同土层深度及不同管距位置的pH值(P0.05)。不同渗灌次数,不同土层深度管中的水溶性盐含量均高于管上和管近,且最大值都出现在1水,较CK分别增加了83.26%和96.34%;不同土层深度,2水的水溶性盐含量平均值都低于1水和3水;不同渗灌次数,不同土层深度,钠离子和钾离子含量的最大值均出现在管中。不同渗灌次数,不同土层深度,管中的水解性氮含量均高于管上和管近,最大值都出现在1水管中,较CK分别增加了112.68%和34.48%。随着渗灌次数的增多,0~10cm土层深度不同管距的速效磷含量均呈现下降趋势,速效钾含量均呈现上升趋势。随着灌水次数的增加,不同土层深度及管位的有机质含量均呈现先上升后下降的趋势,2水显著提高了各位置土壤有机质含量(P0.05)。综上所述,渗灌2次比较好,土壤水溶性盐含量较低且有机质含量较高,利于苜蓿幼苗生长。     

0~4周龄云岭红肉鸡赖氨酸适宜需要量的研究

试验通过分析饲粮不同赖氨酸水平对云岭红肉鸡生长性能、氮沉积率及血液生化指标的影响,确定云岭红肉鸡饲粮赖氨酸的适宜需要量。采用单因素试验设计,将180只健康状况良好、体重相近的1日龄云岭红肉鸡随机分为5个处理组,每组3个重复,每个重复12只鸡。5组试验鸡饲喂的饲粮赖氨酸水平分别为0.85%、0.95%、1.05%、1.15%、1.25%。结果表明:赖氨酸水平对试验鸡的期末体重、全期增重、平均日增重及料重比有显著影响(P<0.05),当赖氨酸水平为1.15%时期末体重最大739.21 g,全期增重最大701.66 g,平均日增重最大24.19 g,料重比最低1.68。不同赖氨酸水平对丝氨酸、甘氨酸和组氨酸的表观代谢率影响显著(P<0.05)。对血清总氨基酸和血清尿素氮影响显著(P<0.05)。研究表明,0~4周龄云岭红肉鸡适宜赖氨酸水平最适量为1.15%。     

子午岭黑山羊与辽宁绒山羊产肉性能、肉品质、肌肉营养成分和脂肪酸含量比较

1990年,甘肃省庆阳市开始引入辽宁绒山羊对子午岭黑山羊进行杂交改良,但目前尚不清楚2个品种在脂肪酸含量、肌肉营养成分等方面的差异,影响了杂交改良效果。试验旨在分析两个绒山羊品种的产肉性能、肉品质、肌肉营养成分和脂肪酸含量差异,为绒山羊的杂交改良提供理论依据。本研究选取相同饲养管理条件下、9月龄的子午岭黑山羊和辽宁绒山羊公羊各5只,测定其屠宰性能以及背最长肌、前腿肌和后腿肌处的肉品质、脂肪酸含量和肌肉营养成分。结果表明：子午岭黑山羊的胴体重、屠宰率、净肉重、净肉率、眼肌面积、GR值、剪切力和滴水损失低于辽宁绒山羊（P<0.05）,但其肌肉的平均亮度值、色度值、pH₁和pH₂₄高于辽宁绒山羊（P<0.01）。营养成分测定结果表明,子午岭黑山羊肌肉的水分和粗灰分含量高于辽宁绒山羊（P<0.05）,但肌内脂肪和粗蛋白含量低于辽宁绒山羊。在2个山羊品种的肌肉中均检测到11 种饱和脂肪酸（SFA,以棕榈酸和硬脂酸为主）、10种多不饱和脂肪酸（PUFA,以亚油酸和顺-11,14-二十碳二烯酸为主）和6种单不饱和脂肪酸（MUFA,以油酸为主）,子午岭黑山羊肉中的SFA、PUFA、n-3 PUFA、n-6 PUFA含量和PUFA/SFA值均高于辽宁绒山羊（P<0.01）,但MUFA含量低于辽宁绒山羊（P<0.01）。结果表明,辽宁绒山羊有更高的产肉力,但子午岭黑山羊肌肉品质和营养成分更佳,脂肪酸组成和含量更符合人类健康膳食标准。     

外源ACC通过激活乙烯合成及信号转导诱导小豆抗锈性

大量研究表明, 乙烯可激发植物对死体营养型真菌的抗性, 但我们前期研究发现, 乙烯合成前体ACC可提高小豆对活体营养型真菌——锈菌的抗性, 为初步明确其机制, 本研究分析了ACC处理对小豆乙烯合成及信号转导的影响, 结果表明, ACC处理显著提高了乙烯合成基因VaACS1及信号通路关键基因VaEIN2?VaEIN3?VaERF5的表达水平?此外, ACC处理后再接种锈菌, 小豆锈病的发病程度显著降低?对接种锈菌后不同时间VaPR2和VaPR4的表达分析表明, 相比ACC处理后不接种对照, ACC处理后再接种锈菌的处理, 接种后1~5 d这两个基因表达量显著升高; 与水处理不接种锈菌相比, 水处理接种锈菌5~8 d后VaPR2和VaPR4的表达量虽显著上调, 但应答时间较ACC处理滞后, 且总体表达水平低, 表明ACC激活乙烯通路进而诱导防卫反应基因上调表达是其诱导小豆抗锈性的关键?