转基因水稻对稻瘟病的抗性研究

 采用苗期初筛、复筛、抗谱测定和田间自然诱发试验等不同鉴定方法,对经分子检测证明已整合有碱性几丁质酶基因和β-1,3-葡聚精酶基因的22个转化系的转基因水稻植株进行稻瘟病抗性鉴定研究,筛选出对稻瘟病的抗性比原种对照七丝软占有明显提高的一系列转基因水稻品系,其中表现高抗的有来自F4-9转化株系的7个品系。高抗材料的R7代品系,经室内抗谱测定及田间病圃试验结果,仍然表现高抗稻瘟病。本研究通过转基因技术,成功地将优质感病品种改良成高抗品系,研究结果证明了利用基因工程手段培育抗病水稻新品种是一个非常有希望的育种途径。     

利用远缘杂交创造核果类果树新种质的研究

以桃、杏、李、樱桃、杏梅等果树的 1 4个品种为亲本 ,3年来共进行了 80余个组合的远缘杂交试验 ,探讨了核果类果树远缘杂交的亲和性 ,并对远缘杂种幼胚进行了胚抢救。结果表明 ,铃铛花期授粉的坐果率显著高于初花期授粉 ;同一杂交组合 ,正反交坐果率差异显著 ,母本对远缘杂交亲和性影响很大 ,选择自交亲和或自然坐果率高的种或品种做母本容易克服远缘杂交的不亲和性 ;适宜场强的静电场、He -Ne激光处理及60 Coγ射线与He -Ne激光联合处理花粉 ,均能显著提高花粉离体萌芽率 ,用上述处理的花粉进行的远缘杂交坐果率也明显高于对照 ;60 Coγ射线单独处理则降低了花粉离体萌芽率 ,远缘杂交的坐果率也低于对照 ;对远缘杂种幼胚及时地进行胚抢救 ,并诱导形成多丛芽 ,是克服核果类果树远缘杂种不育性的有效方法。研究并筛选出了李、樱桃胚萌发与生长、多丛芽诱导与增殖以及生根培养等最佳培养基配方。目前已将欧洲甜樱桃×中国樱桃、大石早生李×泰安巴旦水杏及凯特杏×总统李等一批核果类果树远缘杂种定植于露地 ,其中欧洲甜樱桃与中国樱桃的种间杂种是国内外首次获得     

三尖杉枝叶粉末防治花生根结线虫病

采用盆栽及大田小区试验,研究了三尖杉Cephalotaxus fortunei枝叶干粉末对花生根结线虫病的防治效果.盆栽试验结果表明,每盆(2kg土)施用三尖杉枝叶干粉末10、15及20 g三种处理,与对照组相比,初侵染相应推迟4、6、10天,并能减少侵染量,降低根结增长率,显著减轻花生根结线虫病病情.大田小区试验结果与盆栽试验结果相似,每小区(5m2)沟施三尖杉枝叶干粉末120、80、50 g,处理后34及53天的2次平均防治效果分别为89%、82%及50%,施药对照10%益舒宝颗粒剂25g处理(相当于45kg/hm2)为74%.综合使用剂量及花生生长情况,建议沟施150kg/hm2三尖杉枝叶干粉为宜.     

额济纳旗浅层地下水环境研究

通过 2 0 0 3年 4- 5月在黑河下游额济纳分两条路线 (a -a’和b -b’)采集水样 ,分析了额济纳现状水资源特征。远离河道区域水化学类型有HCO3·SO4-Na、Cl·SO4-Na·Ca、HCO3·Cl-Na·Ca和SO4·Cl-Na ;在河道附近或河道地区水化学类型主要是SO4·HCO3-Na ,类型单一 ;研究区矿化度和各离子含量随距离补给源的远近增减而升降 ,表明它们主要依赖于上游补给水量 ;同时 ,额济纳地处干旱区 ,降雨稀少 ,该区植被的生长发育主要依靠浅层地下水 ,地下水环境的改变直接导致了区域生态的变化。     

Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.     

The ex vivo expansion characteristic of endothelial progenitor cells

AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34⁺ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34⁺ and CD34^- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34⁺ and CD34^-,total MNC led to a significant increase in the expansion of CD34⁺ cells compared with CD34 enrichment (P＜0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P＞0.05). These differentiated EPCs were stained positive for CD34⁺, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34⁺ and AC133⁺cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34⁺ and CD34^- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs.     

Radioligand binding assay of insulin receptor in rabbit kidney during ischemic and reperfusion

AIM: To observe the change of insulin receptor in rabbit kidney with acute ischemic-reperfusion injury. METHODS: 15 Japanese white rabbits were allocated randomly into control group, ischemic-reperfusion group(IR group). IR group received clamping for 1 h followed by 2 h or 48 h of reperfusion. At 2 h or 48 h after reperfusion, glucose and insulin in serum were determined. Insulin receptor in renal tissue was analyzed by radioligand binging assay(BAD). RESULTS: The level of serum glucose increased after 2 h reperfusion in 2 groups, but in IR group the value increased much more higher than those in control groups(P＜0.05). Plasma insulin of IR group was significantly higher than that in control after 2 h reperfusion(P＜0.05). Scatchard analysis of data resulted in curvilinear profiles, indicating that there are two classes of receptors with different affinity or the presence of a single class of receptors with a negative cooperative hormone-receptor interaction. Data analyzed for a two-site model showed that the values of Bmax₁(high affinity site), Bmax₂(low affinity site) and Kd₁, Kd₂ were significantly lower than that of control (P＜0.05) after 2 h perfusion. 48 h after IR there was no difference of Bmax₁, Bmax₂, and Kd₁ between 2 groups,but Kd₂ of IR group was higher than that of control (P＜0.05). CONCLUSIONS: The results indicate that the effect of intrinsic insulin decreases in the progress of the renal ischemic-reperfusion. The resulting high serum glucose may aggravate renal injury in the progress of ischemic-reperfusion.     

Effect of atrial natriuretic peptide on acute lung injury induced by lipopolysaccharide

AIM: To investigate effect of atrial natriuretic peptide (ANP) on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rat. METHODS: Mean arterial blood pressure (MAP) was recorded with model 6280 physiology intelligentialize grapher, nitric oxide (NO) and endothelin (ET) concentrations in plasma were measured after lipopolysaccharide (LPS) or following LPS ,ANP was injected into vein in rats. After experiment,lung water as well as pulmonary histopathological changes was measured and observed, respectively. RESULTS: Administration of LPS elicited a persistence decrease in MAP (8.1 kPa±2.6 kPa,at 4 h,P<0.01 vs control); NO and ET concentration in plasma was evident higher than that in control group, respectively (P<0.01); Wet-dry ratio of lung was higher than that in control group (5.15±0.43,at 4 h) (P<0.05); Alveolus detelectasis was observed and pulmonary mesenchyme was thicker than that in control group. No erythrocyte and leukocytes in alveolus,which show an interstitial pulmonary edema, was observed in LPS+ANP group, ANP maintained MAP at higher levels (13.35 kPa±2.93 kPa, at 4 h, P<0.05 vs LPS) after an transient decline when LPS was injected; NO and ET concentration of plasma had all significantly decrease, respectively (P<0.05 vs LPS, at 4 h); Wet-dry ratio of lung was lower than LPS group (4.57±0.35, P<0.05). Compared with control group the ratio was not evident difference (P>0.05); The histopathological of lung displayed markedly improved. CONCLUSION: ANP attenuates ALI induced by LPS in the rat. The effect of ANP may be via decreasing secretion of ET,NO and regulation arterial blood pressure.     

规模化猪场大肠杆菌的耐药性监测及血清流行病学调查

为了调查猪致病性大肠杆菌的耐药性及流行血清型 ,从湖北、河南、江西、安徽、浙江等省的 33个猪场采集 32 1份仔猪黄、白痢病料进行细菌的分离和生化鉴定 ,结果分离到大肠杆菌 30 2株 ,其中致病性大肠杆菌 2 76株。对 112株致病性大肠杆菌进行了 15种抗生素敏感性试验 ,发现分离菌株对 15种药物均有不同程度的耐药性 :青霉素 G对其完全没有抑制作用 ;先锋霉素 抑制作用最强 (97.3% ) ,其次为呋喃妥因 (78.6 % )、痢特灵 (71.4 % )、环丙沙星(6 8.8% )、诺氟沙星 (6 5 .1% )。 112株菌中 ,有 4 7种耐药谱型 ,多数为 6耐以上的菌株 (80株 ) ,占供试菌株的 71.4 %。应用微量平板凝集试验 ,对分离的 2 76株致病菌进行了血清型鉴定 ,鉴定共有 72种血清型 ,其中优势血清型 10种 ,占能定型分离致病菌株的 5 3.7%。     

3周龄断奶仔猪胰酶发育规律的研究

以 3周龄断奶仔猪为试验材料 ,研究了断奶对仔猪胰酶活性的影响。试验表明 ,3周龄断奶可以显著降低仔猪胰脏中胰淀粉酶、胰蛋白酶、胰糜蛋白酶和胰脂肪酶的活性 ,且在断奶后 2周继续维持断奶时水平。断奶降低了小肠内容物中胰淀粉酶和胰脂肪酶的活性 ,但空肠和回肠胰蛋白酶、胰糜蛋白酶的活性在断奶后并未下降。