以Taichung29为背景的小麦抗条锈病近等基因系转育进展

自1988年起,系统开展了以Taichung29为背景的小麦抗条锈病近等基因系转育及其基础性研究。采用回交法和系谱法相结合的转育方法,以春性品种Taichung29为轮回亲本作母本,分别与25个抗性供体即中国小麦条锈病菌鉴别寄主和国际上重要的抗条锈基因载体品种杂交、回交和自交。通过基因推导分析、单体分析和SSR标记技术检测目的基因,选育抗条锈近等基因系,现已获得重要进展,成功选育出8个以Taichung29为背景的抗条锈病单基因近等基因系,即Taichung29*6／Yr1、Taichung29*6／Yr2、Taichung29*6／Yr5、Taichung29*6／Yr7、Taichung29*6/Yr9、Taichung29*6／Yr10、Taichung29*6/YrSpP、Taichung29*6/YrKy2。另有9个组合转育获得343个抗性稳定株系,正检测其目的基因,3个组合转至BC₆F₃,自交纯合筛选抗性稳定株系,5个组合转至BC₅,继续回交转育。     

Bt玉米对中红侧沟茧蜂发育的间接影响及室内行为研究

利用Y-型嗅觉仪研究了中红侧沟茧蜂(Microplitis Mediator Haliday)对Bt玉米、常规玉米、粘虫幼虫和其虫粪及虫害苗挥发物的行为反应,同时研究了Bt玉米对中红侧沟茧蜂发育的影响。结果表明,两种玉米健康植株的挥发物对中红侧沟茧蜂均有引诱作用;中红侧沟茧蜂对两种玉米的健康植株及机械损伤株挥发物之间的选择性无显著差异;相对于Bt玉米,中红侧沟茧蜂更趋向于选择常规玉米的虫伤苗及玉米-粘虫幼虫-虫粪混合物的挥发物。与对照(寄生于取食常规玉米粘虫的中红侧沟茧蜂)相比,寄生于取食Bt玉米粘虫幼虫的中红侧沟茧蜂幼虫历期延长,出茧率、茧重、羽化率、蜂重均有显著降低,茧历期、蜂历期则差异不显著。     

百合病毒病的发生与症状类型

百合病毒病症状类型可归纳为7种类型,轻花叶型(Mm),重花叶型(Sm),矮化型(Stu),丛簇矮化型(Rstu)。黄化矮化型(Ys),扁茎簇叶型(Fsbl),花变叶型(Phy),其中轻花叶型和重花叶型发生普遍,花变叶型出现较少。病害发生的轻重与种球种龄有关,种龄越大,发病越重。前茬种植百合的地块较种植小麦的地块发病重,低洼积水地发病重。不同百合品种对病毒病的抗性有差异。     

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.     

Protective effect of HSP70 on injuried myocardial cells induced by H2O2

AIM: To investigate the role of heat-shock protein 70(HSP₇₀) in the protection of myocardial cells against ischemic injury.METHODS: Myocardial cells were cultured in vitro. HSP₇₀ was induced by hyperthermia. H₂O₂-injured myocardial cells were divided into different groups. Flow cytometry, DNA Ladder and biochemistry methods were employed to observe the myocardial cells of different groups.RESULTS: Immunohistochemistry showed hyperthermia induced the up-regulation of HSP₇₀ in myocardial cells. Apoptotic rate, activity analysis of cytochrome C and succinic dehydrogenase in H₂O₂-injuried and HSP₇₀-protected groups were obviously different. Electron micrograph shomed hyperthermia alliviated myocardial cell injury induced by H₂O₂. CONCLUSION: HSP₇₀ delays apoptosis and protects against H₂O₂-induced myocardial cell injury.     

Taurine reduced oxygen free radical production induced by homocysteine in electron transport chain and homocysteine inhibites taurine transporter in mitochondria membrane

AIM: To observe effects of homocysteine and antagonized effects of taurine on electronic leakage and free radical production in myocardial mitochondria. METHODS: Myocardial mitochondria of rat heart was isolated, and was broken by supersonic wave to prepare submitochondria. Recombinant of succinic acid cytochrome c reductase was prepared with mitochondria of porcine heart. They were co-incubated with homocysteine and/or taurine with various concentration. The H₂O₂ and O₂^- were determined by chemiluminescence methods. The taurine transporter of heart mitochondria and its propert, and effects of homocysteine on its function were studied with glass filter. RESULTS: Homocysteine stimulated oxygen free radical production in heart mitochondria, submitochondria, and succinic acid cytochrome c in a concentration-dependent manner. Although taurine itself did not affect oxygen free radical production, taurine did inhibit oxygen free radical production in mitochondria, submitochondria and succinic acid cytochrome c in a concentration-dependent manner. Taurine transporters of Na⁺-dependent were existed in mitochondria membrane. Homocysteine inhibited taurine transtport in mitochondria in a concentration-dependent manner. CONCLUSIONS: Taurine inhibited electronic leakage and oxygen free radical production induced by homocysteine in electron transport chain. There were taurine transporters in mitochondria membrane, and transport functions of taurine transporter were inhibited by homocysteine.     

Effect of sodium nitroprusside on activation of nuclear factor κB

AIM: To investigate the effect of sodium nitroprusside (SNP) on activation of nuclear factor κB. METHODS: The techniques of culture of human T lymphocytes, Western blot and RT-PCR were applied. The effects of NO donor sodium nitroprusside (SNP) at different concentrations on mRNA and protein expression of IκB_α in human T lymphocytes at 30 min or 120 min after stimulating with phytohaemagglutinin (PHA-P) were observed. RESULTS: SNP at middle or high concentrations reduced the degradation of κIB_αprotein 30 min after stimulating with PHA-P,and increased the re-expression of κIB_αmRNA 120 min after stimulating with PHA-P significantly.CONCLUSION: The mechanism of inhibitory effect of SNP at middle or high concentrations may be due to the decrease in degradation and the increase in re-synthesis of κIB_α.The regulatory mechanism of SNP at low concentration may not be through κIB_α.     

Expression and subcellular localization of urocortin in syncytiotrophoblast of human term placenta

AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine.     

Gene expression of 5α-reductase (type 2) in reproductive gland of pubertal diabetic rats

AIM: To study the molecular mechanism of reproduction dysfunction in pubertal diabetic rat, the level of 5α-reductase (type 2) mRNA in testis, epididymis and prostate in diabetic rat was detected. METHODS: The gene expression of 5α-reductase (type 2) was detected by Northern blot. RESULTS: In the caput of the epididymis, the expression of 5α-reductase (type 2) mRNA of D and ID groups was less than that of C group. CONCLUSION: The decreased expression of 5α-reductase (type 2) results in the decreased production of dihydrotestosterone, which influences the development and function of reproduction system in pubertal rats.     

Effects of inhibition of Kupffer cell and splenectomy on thioacetamide-induced hepatic injury

AIM: To examine the effects of inhibition of Kupffer cell and splenectomy on intestinal endotoxemia and hepatic injury. METHODS: The hepatic injury model was established by treatment with thioacetamide (TAA). At the same time, inhibition of Kupffer cells by intravenous GdCl₃ and splenectomy were performed. Serum alanine aminotransferase (ALT), TNF-α, endotoxin content and phagocytic index were observed. RESULTS: In the TAA+GdCl₃ group, and TAA+splenectomy group, the endotoxin content was significently higher than that in normal and TAA group (P<0.05). The plasma TNF-α, ALT and total bilirubin were significantly lower than those in TAA group (P<0.05). CONCLUSION: Inhibition of Kupffer cells and splenectomy increase plasma endotoxin level, but decreases the plasma TNF-α levels and alleviates the hepatic injury induced by TAA.