Propanil and swep inhibit 4-coumarate:CoA ligase activity in vitro

4-Coumarate:CoA ligase (4CL, EC 6.2.1.12) in the phenylpropanoid pathway in plants has attracted interest as a novel target for developing effective plant growth inhibitors (PGIs). In a previous study in which the 4CL inhibitory activity of 28 existing herbicides was investigated using an optimized in vitro screening assay, 4CL activity was found to be strongly inhibited by propanil and swep at 100 microM. Here, further experimental evidence is provided to substantiate the previous result. Using 4-coumaric acid as substrate, tobacco 4CL activity was inhibited by propanil or swep in a concentration-dependent manner, with 50% inhibition concentrations (I(50)) of 39.6 and 6 microM respectively. These herbicides also exhibited uncompetitive inhibition towards 4-coumaric acid. Furthermore, 4CLs from several plant species were inhibited by the herbicides within a range from 1 to 50 microM. It is proposed that these herbicides have another site of action as a result of the inhibition of 4CL in the phenylpropanoid pathway, and this enzyme represents a new target site for the development of PGI.     

氮素调控对玉米氮素同化过程及产量的影响

氮素调控措施与作物氮素吸收利用和产量密切相关,但目前关于不同氮素调控措施对玉米主要生育期氮素同化过程的影响仍不清楚。以郑单958为试验材料,设置不施氮肥（CK）、传统施肥（CN）、氮肥+生物炭（SN）和氮肥+硝化抑制剂DMPP（DN）4个处理,分析不同氮素调控对玉米氮素同化过程中铵态氮和硝态氮含量、硝酸还原酶（NR）和谷氨酰胺合成酶（GS）活性、游离氨基酸和可溶性蛋白含量以及氮素利用率和产量的影响。结果表明：DN和SN处理较传统施肥处理均可以提高玉米植株体内硝态氮和铵态氮含量、NR和GS活性;DN和SN处理显著提高灌浆期谷氨酸、游离氨基酸和可溶性蛋白含量;DN处理成熟期籽粒的氮素积累量显著高于SN和CN处理,分别显著增加18.4%和30.0%;DN处理产量最高,SN次之,二者并无显著差异,但相较CN处理分别显著增产1 483.0和1 154.2 kg·hm^-2。两种氮素调控均促进了玉米对氮素的吸收,显著提高氮肥吸收利用率,其中硝化抑制剂处理氮肥吸收利用率最高且显著高于其他处理。综上,生物炭或硝化抑制剂配施氮肥,可以促进玉米氮素同化和转运过程,显著提高玉米产量和氮肥利用效率,综合产量、籽粒氮素积累量和氮肥吸收利用率,硝化抑制剂配施氮肥可作为淮河流域玉米高产高效的栽培措施。     

A Cathepsin B Inhibitor,E-64, Improves the Preimplantation Development of Bovine Somatic Cell Nuclear Transfer Embryos

Bovine somatic cell nuclear transfer (SCNT) is an important and powerful tool for basic research and biomedical and agricultural applications, however, the efficiency of SCNT has remained extremely low. In this study, we investigated the effects of cathepsin B inhibitor (E-64) supplementation of culture medium on in vitro development of bovine SCNT embryos. We initially used three concentrations of E-64 (0.1, 0.5, 1.0 μm), among which 0.5 μm resulted in the highest rate of blastocysts production after in vitro fertilization (IVF), and was therefore used for further experiments. Blastocyst development of SCNT embryos in the E-64 treatment group also increased relative to the control. Moreover, the cryosurvival rates of IVF and SCNT blastocysts were increased in E-64 treatment groups when compared with the control. On the other hand, we found that IVF and SCNT blastocysts derived from E-64-treated groups had increased total cell numbers and decreased apoptotic nuclei. Furthermore, assessment of the expression of apoptosis-related genes (Bax and Bcl-xL) in bovine IVF and SCNT blastocysts treated with E-64 by real-time RT-PCR analysis revealed suppressed expression of the pro-apoptotic gene Bax and stimulated expression of the anti-apoptotic gene Bcl-xL. Taken together, these finding indicate that addition of E-64 to embryo culture medium may have important implications for improving developmental competence and preimplantation quality in bovine IVF and SCNT embryos.     

Effect of idazoxan on permeability of blood-brain barrier and expression of MMP-9/TIMP-1 in mouse experimental autoimmune encephalomyelitis

AIM：To study the effect of idazoxan (IDA) on the permeability of blood-brain barrier (BBB) and the expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in mouse experimental autoimmune encephalomyelitis (EAE).METHODS：Female C57BL/6 mice (n=36) were randomly divided into control group, EAE group and IDA group, with 12 mice in each group. EAE was induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55). IDA (2 mg/kg, ip, bid) was administered for 15 d after immunization. The neurological defects of the mice were observed daily and scored. The pathological changes were observed under microscope with HE staining and LFB myelin staining. The BBB permeability was detected by Evans blue extravasation. The expression of MMP-9 and TIMP-1 in the brain of EAE mice was determined by Western blotting.RESULTS：Compared with EAE group, the score of neurological defects in IDA group was decreased, the inflammation was relieved, the BBB permeability was reduced, and the expression MMP-9 and the ratio of MMP-9/TIMP-1 were decreased (P<0.05).CONCLUSION：The neuroprotective effect of IDA on mouse EAE might be related to the down-regulation of MMP-9 and the ratio of MMP-9/TIMP-1, thus reducing the degradation of BBB and the permeability of BBB, and ameliorating the pathologic process of EAE.     

钙信号抑制剂加剧低钾胁迫对烟草幼苗光合特性及钾吸收的影响

  【目的】  研究低钾胁迫抑制钙信号传导对植株幼苗叶片光合特性及钾吸收的影响,为缓解作物的低钾胁迫提供理论依据。  【方法】  以烟草品种K326为试验材料进行了室内水培试验。烟草幼苗先在营养液正常钾(K+ 5 mmol/L)和缺钾(K+ 0.15 mmol/L)营养液中生长8 天,然后在营养液中分别施加钙通道有机阻断剂Vp、钙通道无机阻断剂LaCl₃和钙离子螯合剂(EGTA)钙信号抑制剂,继续生长4天后取样,测定植株干鲜重、电导率、活性氧含量、叶片抗氧化酶活性、叶片光合特性等指标和植株地上、地下部钾素浓度及钾离子吸收相关基因表达量。  【结果】  1)不添加钙通道抑制剂或螯合剂条件下,低钾胁迫的烟株地上部和根系干重较常钾水平显著降低了59.29%、39.82%,鲜重显著降低了51.58%、48.19%,除水汽压饱和亏外的其余5项光合特性指标显著降低,植株地上部和根系钾含量分别显著降低了39.31%和77.05%,而叶中可溶性蛋白、相对电导率、过氧化氢含量则显著增加,NKT2、NtKC1、NtHAK1、NtHAK5基因的相对表达量分别增加了2.09、3.32、3.23、12.34 倍。2)常钾水平下,3个钙抑制剂处理未明显影响地上部和根系干鲜重,而低钾水平下,钙抑制处理的地上部鲜重和干重显著高于无钙抑制剂对照。两个钾水平下,与各自对照相比,3个钙抑制剂处理的叶片相对电导率、活性氧积累量均显著增加,抗氧化酶SOD和POD活性、叶绿素含量显著减少,光合特性指标(除水汽压饱和亏外)明显降低。3)两个钾水平下,3个钙信号抑制剂处理的烟株地上部和根系钾含量均显著低于其对照,低钾胁迫下平均降低幅度分别达到25.54%和53.36%。低钾胁迫下,钾离子通道基因NKT2、NtKC1和转运蛋白基因NtHAK1、NtHAK5在根系中的相对表达量分别平均降低了60.60%、50.62%、38.61%、69.79%,钙通道相关基因NtCNGC1和NtCNGC11在根中的相对表达量分别平均降低了95.36%和87.04%。  【结论】  低钾胁迫明显影响烟株幼苗生长、光合作用及对钾素的吸收和积累。低钾水平下,外施钙通道抑制剂或螯合剂可进一步降低烟草叶片抗氧化酶活性,减弱烟株叶片光合作用,并抑制根中钾吸收相关基因的表达和烟株对钾素的吸收。     

HBV衣壳蛋白装配抑制剂的研究进展

乙型肝炎病毒(HBV)是一种常见的嗜肝性病原体,全球约有4亿人感染,带来严重的社会问题和经济负担。虽然目前已有预防和治疗HBV的药物,但由于耐药性等问题不能彻底清除病毒感染,新药开发依然迫在眉睫。介绍了HBV的病毒结构、复制和感染过程,对现有治疗药物的优缺点进行了总结。对以衣壳蛋白装配为靶点的新药研究进展进行综述,旨在为今后开发抗HBV的新药提供参考。     

脲酶测定法判定豆浆抗营养因子热失活情况研究

大豆抗营养因子的主要成分是胰蛋白酶抑制剂,在大豆制品加工过程中,通常采用高温加热钝化的方法使胰蛋白酶抑制剂失去活性。豆浆是人们日常生活中喜欢的饮品之一,因为脲酶测定方法比较简单,目前多以加热钝化后豆浆脲酶活性来表示抗营养因子的去除情况。本研究通过测定豆浆中胰蛋白酶抑制剂和脲酶的活性研究豆浆加热过程中二者失活是否具有一致性,判定以脲酶活性来体现豆浆安全的科学性,同时通过豆浆加热程度与胰蛋白酶抑制剂失活情况的关系,对家庭豆浆制作提供一种安全的加热模式。