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.     

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。     

减氮配施硝化抑制剂与菌剂对温室黄瓜产量品质和土壤氮素损失的影响

通过探究减氮配施硝化抑制剂DMPP与微生物菌剂及二者联合施用对温室黄瓜土壤氮素各主要途径损失及黄瓜对氮素吸收利用的影响,并结合黄瓜产量和品质,旨在筛选出温室黄瓜生产的适宜氮素损失调控措施。以黄瓜品种津绿20-10为试验材料进行田间小区试验,设置6个处理,分别为不施氮对照(CK)、常规施氮(CN)、减氮(RN)、减氮+DMPP(RND)、减氮+微生物菌剂(RNM)、减氮+DMPP+微生物菌剂(RND+M)。监测分析了土壤氧化亚氮(N₂O)排放、氨(NH₃)挥发和土壤剖面硝态氮(NO₃^--N)累积量,以及黄瓜对氮素的吸收利用、产量和品质指标。结果表明:(1)与CN相比,RN、RND、RNM和RND+M能够促进黄瓜对氮素的吸收和利用,提高氮素利用率。等氮条件下,RND+M可使黄瓜地上部植株氮素总吸收量增加18.93%,尤其是氮肥表观利用率(REN)和农学效率(AEN),分别达到25.30%和41.16 kg/kg(p<0.05),表现出明显的正协同效应,且优于硝化抑制剂或菌剂单施效果。(2)RN、RND、RNM和RND+M较CN可使土壤N₂O排放显著降低26.38%~41.45%、NH₃挥发明显减少28.82%~37.70%,0—120 cm土壤剖面NO₃^--N累积显著降低13.07%~62.32%;等氮条件下,RNM处理对土壤N₂O排放和NH₃挥发影响不大,但能显著降低90—120 cm土层NO₃^--N累积量,较RN降低27.35%。RND和RND+M可使N₂O排放分别降低20.11%和20.47%,0—120 cm土壤剖面NO₃^--N累积量分别降低30.06%和24.70%,减少氮素在土壤中的累积和淋失风险,但增加NH₃挥发风险(p＞0.05),总体表现为RND≈RND+M≥RNM≈RN。(3)RND+M处理产量为70.32 t/hm²,节本增收较RN增加5 150元/hm²,且其在提高黄瓜果实品质方面效果较明显,可溶性蛋白含量较RN及RNM处理分别提高16.36%与4.01%。综合经济效益和环境效益,尤其是土壤可持续发展角度考虑,试验条件下,追施氮素316 kg/hm²,同时配施2%纯氮量的DMPP与75 L/hm²菌剂,是实现温室黄瓜增产提质、绿色高质量发展的适宜氮素损失调控措施。     

抑制膳食相关酶活性的红曲霉菌株的选育

为弄清红曲霉在降脂减肥方面的作用,进一步开发红曲产物类药物应用于市场,以产红曲红色素的红曲霉菌株为出发菌株,经60Co诱变,对抑制与膳食相关的α-淀粉酶、胰蛋白酶和脂肪酶3种酶活性的红曲霉菌株进行了选育。结果表明:选育出1株能显著抑制3种与膳食相关酶的突变菌株M3-19,菌株M3-19液体发酵产物对α-淀粉酶、胰蛋白酶、脂肪酶有明显的抑制作用,抑制率分别为55.83%、25.4%和31.5%。经传代10代遗传稳定。     

食品防霉剂富马酸二甲酯的合成

以四氯化锡为催化剂,马来酸酐和甲醇为原料,采用"一锅法"催化合成富马酸二甲酯。考察了反应时间、醇酸摩尔比、催化剂的用量对产率的影响,并进行了正交实验设计。结果表明:醇酸摩尔比对产率的影响高度显著,当甲醇与马来酸酐的摩尔比为15∶1,四氯化锡质量分数0.8g,回流反应时间2.5h,产率达88.72%。通过熔点测试和红外光谱分析,确定合成产物即是富马酸二甲酯。     

4种荚蒾属植物种子内源抑制物存在特征初探

通过荚蒾种子浸提液对小白菜种子萌发的抑制作用,分析探讨荚蒾种子内源抑制物主要存在部位及其可溶性化学特征,为荚蒾属植物种子破除休眠提供科学依据。实验结果表明:4种荚蒾种子水和甲醇浸提液对小白菜种子萌发和胚根生长都有抑制作用,种皮水溶性抑制物活性:蒙古荚蒾朝鲜荚蒾暖木条荚蒾鸡树条荚蒾,胚乳水溶性抑制物活性:蒙古荚蒾鸡树条荚蒾暖木条荚蒾朝鲜荚蒾;种皮脂溶性抑制物活性:暖木条荚蒾蒙古荚蒾朝鲜荚蒾鸡树条荚蒾,胚乳脂溶性抑制物活性:鸡树条荚蒾朝鲜荚蒾蒙古荚蒾暖木条荚蒾。鸡树条荚蒾、朝鲜荚蒾、暖木条荚蒾内源抑制物主要是脂溶性物质,前两者较集中分布于胚乳,后者多存在于种皮;蒙古荚蒾以胚乳中水溶性抑制物为主。     

水稻黑条矮缩病毒S6片段的序列多样性与进化分析

<正>0 引言植物RNA病毒在进化中具有快速变异的能力,有利于病毒扩大寄主范围甚至进化为新病毒[1]。突变和重组是植物RNA病毒变异和遗传多样性的共同进化因素,是群体基因变异的最初来源[2]。由灰飞虱传播的水稻黑条矮缩病毒(Rice black streaked dwarf virus, RBSDV) 属于呼肠孤病毒科(Reoviridae)斐济病毒属(Fijivirus),能引发玉米粗缩病和水稻黑条矮缩病。