MicroRNA-145 inhibits epithelial-mesenchymal transition in human renal cancer A-498 cells by ADAM28

AIM: To investigate the inhibitory effect of microRNA-145 (miR-145) on epithelial-mesenchymal transition (EMT) in renal cancer A-498 cells. METHODS: The A-498 cells were transfected with miR-145 mimics (M145) and mimic negative control(MNC), which served as M145 group and MNC group, respectively. Mock control (MC) group was set up using untreated A-498 cells. The expression level of miR-145 in each group was detected by RT-qPCR. Transwell assay was used to detect the invasion ability of the cells. The protein expression of vimentin, E-cadherin and ADAM28 was determined by Western blot. Bioinformatic method was used to predict the target genes of miR-145. Antagonistic effect of ADAM28 over-expression on the inhibition of EMT by miR-145 was detected by Western blot. The relationship between miR-145 and ADAM28 was analyzed by dual-luciferase reporter assay. RESULTS: The expression level of miR-145 in M145 group was significantly up-regulated than that in MC group (P<0.05). The number of invasive cells in M145 group was 12.78±3.37, which was significantly lower than that in MC group (P<0.05). ADAM28 may be the target gene of miR-145. Compared with MC group, the protein expression of vimentin and ADAM28 in M145 group was significantly decreased (P<0.05), while the protein expression of E-cadherin was significantly increased (P<0.05).After ADAM28 over-expression, the protein expression of vimentin in the A-498 cells of M145 group was significantly increased (P<0.05), and the protein expression of E-cadherin was significantly decreased (P<0.05). The results of dual-lucife-irasei reporter assay showed that ADAM28 was a downstream target gene of miR-145. CONCLUSION: miR-145 may inhibit the expression of EMT-related proteins through the downstream target gene ADAM28 and inhibit the EMT process of renal cancer A-498 cells.     

Leaf carbohydrates assimilation and metabolism affect seed yield of rapeseed with different waterlogging tolerance under the interactive effects of nitrogen and waterlogging

Waterlogging is a main factor causing rapeseed (Brassica napus L.) yield loss, and reasonable nitrogen (N) applications can compensate for this loss. To investigate the effects of N rates on seed yield of waterlogged rapeseed, the waterlogging-tolerant rapeseed variety ZS 9 and sensitive variety GH01 were waterlogged for 0 and 10 days with five leaves at the seedling stage under four N rates (0, 90, 180 and 270 kg/ha). Waterlogging significantly decreased seed yield, while N application can alleviate the yield loss. The yield decrease rate of waterlogged GH01 was greater than that of ZS 9 under the same N rate. During the seedling and bolting stage, the leaf photosynthetic rate (Pn) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity increased, while activities of adenosine diphosphate glucose pyrophosphorylase (AGPase), sucrose synthase (SuSy) and sucrose phosphate synthase (SPS) decreased with more N under the same watering conditions. Compared to the plants without waterlogging, the leaf Pn and Rubisco activity, starch and sucrose contents of waterlogged rapeseed decreased at the two stages; activities of AGPase, SuSy and SPS of waterlogged rapeseed decreased at the seedling while increased at the bolting stage for both the two varieties. At the flowering stage, the Pn, the activities of Rubisco, AGPase, SuSy, SPS and contents of sucrose, starch increased with more N application for both ZS 9 and GH01. Compared to the plants without waterlogging, the Pn and Rubisco activity for waterlogged plants of the two varieties increased; the waterlogged plants of tolerant variety had higher activities of AGPase, SuSy and SPS, while those of sensitive variety was significantly lower. However, the decreased starch and sucrose content were found in both tolerant and sensitive varieties. The activities of AGPase, SuSy and SPS at flowering were highly positively correlated with yield under the interactive effects of N and waterlogging. These results suggested that the flowering stage is the most important stage that N had the positive regulation on waterlogged rapeseed growth. The carbohydrates translocation from leaves to seeds of the tolerant variety were enhanced after waterlogging, while that of the sensitive variety was still inhibited. This was the main reason for the difference in yield between the two waterlogged varieties.     

利用酵母双杂交系统筛选与草莓镶脉病毒移动蛋白P1互作的寄主因子

 构建感染草莓镶脉病毒(SVBV)森林草莓的酵母cDNA文库,利用酵母双杂交系统,筛选出与SVBV P1蛋白互作的15种寄主因子。生物信息学分析发现,这15种寄主因子参与茉莉酸途径、泛素化、光合作用、抗病抗逆、蛋白修饰、蛋白运输和氧化还原等多种生物过程。另外,这些寄主因子还具有其他分子功能,包括氧化还原酶活性、蛋白二硫化物异构酶活性和金属离子结合活性等。本研究初步探讨了P1与寄主因子的互作机理,为揭示SVBV侵染森林草莓以及SVBV在寄主中扩展的分子机制提供理论依据。     

Effect of dietary calcium or phosphorus deficiency on bone development and related calcium or phosphorus metabolic utilization parameters of broilers from 22 to 42 days of age

This experiment was conducted to investigate the effect of dietary calcium (Ca) or phosphorus (P) deficiency on bone development and related Ca or P metabolic utilization parameters of broilers from 22 to 42 days of age based on our previous study, which indicated that dietary Ca or P deficiency impaired the bone development by regulating related Ca or P metabolic utilization parameters of broilers from 1 to 21 days of age. A total of 504 one-day-old Arbor Acres male broilers were randomly assigned to 1 of 4 treatments with 7 replicates in a completely randomized design, and fed the normal control and Ca- or P-deficient diets from 1 to 21 days of age. At 22 days of age, the broilers were further fed the normal control diet (0.90% Ca+0.35% non-phytate P (NPP)), the P-deficient diet (0.90% Ca+0.18% NPP), the Ca-deficient diet (0.30% Ca+0.35% NPP) or the Ca and P-deficient diet (0.30% Ca+0.18% NPP), respectively. The results showed that dietary Ca or P deficiency decreased (P<0.05) tibia bone mineral density (BMD), bone breaking strength (BBS), ash content, tibia ash Ca content and serum P content on days 28 and 42, but increased (P<0.05) tibia alkaline phosphatase (ALP) activity of broilers on day 42 compared with the control group. Furthermore, the broilers fed the P-deficient diet had the lowest (P<0.05) tibia BMD, BBS, ash content, serum P content and the highest (P<0.05) serum Ca content on day 28 compared with those fed the Ca-deficient or Ca and P-deficient diets. The results from the present study indicated that the bone development and related Ca or P metabolic utilization parameters of broilers were the most sensitive to dietary P deficiency, followed by dietary Ca deficiency or Ca and P-deficiency; dietary Ca or P deficiency impaired the bone development possibly by regulating serum Ca and P contents as well as tibia Ca content and ALP activity of broilers from 22 to 42 days of age.     

Protective efficacy of an H5/H7 trivalent inactivated vaccine produced from Re-11, Re-12, and H7-Re2 strains against challenge with different H5 and H7 viruses in chickens

We developed an H5/H7 trivalent inactivated vaccine by using Re-11, Re-12, and H7-Re2 vaccine seed viruses, which were generated by reverse genetics and derived their HA genes from A/duck/Guizhou/S4184/2017(H5 N6)(DK/GZ/S4184/17)(a clade 2.3.4.4 d virus), A/chicken/Liaoning/SD007/2017(H5 N1)(CK/LN/SD007/17)(a clade 2.3.2.1 d virus), and A/chicken/Guangxi/SD098/2017(H7 N9)(CK/GX/SD098/17), respectively. The protective efficacy of this novel vaccine and that of the recently used H5/H7 bivalent inactivated vaccine against different H5 and H7 N9 viruses was evaluated in chickens. We found that the H5/H7 bivalent vaccine provided solid protection against the H7 N9 virus CK/GX/SD098/17, but only 50–60% protection against different H5 viruses. In contrast, the novel H5/H7 trivalent vaccine provided complete protection against the H5 and H7 viruses tested. Our study underscores the importance of timely updating of vaccines for avian influenza control.     

文心兰高效再生体系的建立

[目的]建立文心兰高效再生体系,为利用转基因技术进行品种改良提供科学依据.[方法]以文心兰柠檬绿的无菌苗叶片为材料,探讨不同植物生长调节剂配比对其原球茎诱导的影响,开展增殖、生根及移栽研究,建立其高效稳定的再生体系.[结果]影响文心兰原球茎诱导的3种植物生长调节剂主次排序为萘乙酸(NAA)>噻苯隆(TDZ)>6-苄氨基嘌呤(6-BA),在1/2MS培养基中添加0.5 mg/L NAA、0.5 mg/L 6-BA和0.3 mg/L TDZ的诱导率最高,达48.0%.培养基中活性炭含量及碳水化合物种类对文心兰原球茎增殖的影响存在明显差异,不同活性炭含量处理间的原球茎增殖系数存在显著差异(P<0.05,下同),添加1.0 g/L活性炭较添加0.5 g/L活性炭的增殖系数显著升高,不同碳水化合物种类间的原球茎增殖系数排序为海藻糖>麦芽糖>蔗糖,最佳增殖和分化培养基为1/2MS+4.0 mg/L 6-BA+0.4 mg/L NAA+1.0 g/L活性炭+20 g/L海藻糖.添加10%椰子水的文心兰幼苗生根数显著高于添加10%香蕉汁和10%苹果汁,每株平均生根量接近4.00条,即生根效果最佳的培养基为1/2MS+0.8 mg/L 6-BA+0.2 mg/L NAA+10%椰子水.移栽过程中根部用水苔包裹后再移至椰糠中,保证光照、水分适宜,适当添加缓释肥,移栽成活率可达100%.[结论]利用海藻糖或麦芽糖作为碳源的文心兰原球茎增殖效果较优,生根数和生根质量优势明显;不同植物生长调节剂配比、碳源和有机物的添加对文心兰组织培养具有明显的促进效果.     

5种蔷薇科树种多酚氧化酶比较基因组学分析

蔷薇科植物中有多种具有重要价值的水果,如苹果、梨、桃、草莓和黑树莓等。这些水果普遍容易发生由多酚氧化酶(PPO)介导的酶促褐变而导致重大经济损失,从全基因组角度分析比较PPO基因家族,有助于加深对PPO基因家族和功能的认识。采用比较基因组学的方法,对这5种植物的PPO基因家族进行了基因鉴定、染色体定位、编码蛋白的亚细胞定位、内含子统计分析、基因系统进化、基因倍增与丢失等特征分析。从这5种植物中,共计鉴定出了42个PPO基因,其中6个基因被认定是假基因;绝大多数基因编码的蛋白定位于叶绿体,2个定位于线粒体,3个是分泌型蛋白;PPO基因在染色体上有串联重复和散布两种形式;9个基因具有内含子,聚类结果显示可以将它们分为6个类型,每个基因类型在进化过程中都发生过基因倍增和丢失;同型基因内含子的位置和大小具有相似性。这些结果揭示蔷薇科植物PPO基因内含子是伴随着基因倍增产生的,基因倍增也是推动PPO基因多样化的重要动力,PPO基因倍增和丢失差异导致PPO基因数量在不同物种之间产生差异。     

新疆生产建设兵团第八师农田残膜含量及在土壤中分布情况

在新疆生产建设兵团第八师农田进行了残膜样品采集，并对不同采样深度下不同面积的农田残膜含量及数量进行统计分析，获取了土壤中残膜的分布情况，旨在为残膜回收机械的设计以及残膜回收策略的制定提供参考。      

60Co-γ射线辐照处理对4种药用植物种子萌发的影响

用不同剂量60Co-γ射线对益母草、蒲公英、车前、紫苏种子进行辐照处理,测定这4种药用植物种子的发芽率、发芽势、发芽指数等指标,了解60Co-γ射线辐照对其种子萌发的影响,为这4种药用植物的新品种选育提供参考。结果表明,60Co-γ射线对4种药用植物种子具有较强的诱发突变能力,辐照剂量与种子发芽率、发芽势、发芽指数、活力指数、苗高、苗鲜重均呈负相关,其中与苗高的相关性达极显著水平。益母草和蒲公英种子的半致死剂量大于400 Gy,车前种子的半致死剂量在300~400 Gy之间,紫苏种子的半致死剂量在50~100 Gy之间。