生物炭对Bt Cry1Ac蛋白抗紫外能力影响

为了研究生物炭对紫外线的防护作用，以豆壳烧制的生物炭作为载体，研究生物炭对Bt Cry1Ac蛋白的吸附行为以及生物炭对Cry1Ac蛋白的紫外保护作用。使用扫描电子显微镜、透射电子显微镜、X-射线粉末衍射以及傅立叶红外光谱等手段对生物炭的形貌和结构进行表征。结果表明，生物炭是典型的多孔结构材料，表面具有丰富的官能团。Cry1Ac蛋白与生物炭吸附平衡时间为50 min，最合适的吸附浓度比（生物炭：蛋白）为1：100，二者吸附符合准二级动力学模型和Langmuir模型。在UVB紫外照射4 h后，生物炭与Cry1Ac蛋白复合物对棉铃虫的生物活性是单纯蛋白的4.93倍，显示生物炭具有较好的紫外抵抗效果。研究结果初步表明，制备得到的生物炭能够显著提高Cry1Ac蛋白的抗紫外能力，为后续研发耐受紫外线的农药剂型提供新材料。     

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.     

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.     

云南怒江傈僳族自治州古树名木资源现状调查

对怒江傈僳族自治州人为活动区域古树名木资源现状进行调查。结果表明:共记录到古树名木496株,涉及36科54属76种,其中有8种国家重点保护野生植物,分别为澜沧黄杉、秃杉、南方红豆杉、云南榧树、千果榄仁、红椿、光叶珙桐和香果树;怒江州古树名木主要分布在乡村街道及远郊野外,占总数的98.8%,63.6%的古树生长在海拔1500~1999 m;古树名木的生长环境及长势总体较好,但保护力度相对薄弱,应采取相关措施加强保护和管理。     

硅藻土吸附固定化Bacillus sp.DL-2胞内蛋白酶拆分制备(S)-乙酸苏合香酯

[目的](S)-乙酸苏合香酯是手性药物合成的关键手性砌块,其在多种手性化合物的合成及医药、香料的工业生产中具有重要的作用.传统的化学合成手性化合物的方法需要有毒有机溶剂及重金属的参与,对环境及人类具有严重的危害,同时得到的手性化合物的光学纯度较低.与传统的化学合成相比,酶法选择性拆分消旋体化合物,具有高立体专一性和区域选择性、副反应少、产率高、产物光学纯度好以及反应条件温和的优点,是一种被广泛认可的拆分方法.实验表明,Bacillus sp.DL-2胞内蛋白游离酶可拆分制备(S)-乙酸苏合香酯,然而游离酶不易回收且稳定性差,将Bacillus sp.DL-2胞内蛋白游离酶制备成固定化制剂,克服了游离酶对环境敏感,稳定性不高的缺点,为工业化生产(S)-乙酸苏合香酯奠定了基础.[方法]酶的固定化方法主要有物理吸附法、离子结合法、共价结合法、交联法和包埋法.以吸附法固定酶,酶的构象很少改变,因此,酶的催化活力损失较少;且吸附法固定化操作过程简单,因此吸附法在经济上是最具吸引力的固定化方法.硅藻土由硅藻的细胞壁沉积而成,硅藻土表面的多孔性与负电性使其呈现明显表面吸附性,因而被常用做吸附载体.以硅藻土作为固定化载体,对Bacillus sp.DL-2胞内蛋白酶进行固定化,用以拆分制备高光学纯的(S)-乙酸苏合香酯.利用单因素实验优化,确定制备固定化酶的最佳固定化条件及制备(S)-乙酸苏合香酯的最佳拆分条件,并测定了制备的固定化酶对金属离子的敏感度及储存稳定性.[结果]最佳固定化条件为:温度40℃,pH为7,固定化时间10 h,载体添加量100 g/L.在最佳固定化条件下制备了固定化酶,优化确定了制备(S)-乙酸苏合香酯的最佳拆分条件为:固定化酶用量为160 mg/mL,拆分时间7 h,拆分温度30℃,缓冲液pH为7.[结论]在最佳固定化及拆分条件下,制备的(S)-乙酸苏合香酯e.e.值可达到96.8％,转化率为73.9％,且固定化酶对金属离子敏感度较低,对反应环境要求较低,适用于工业化生产.固定化酶在4℃条件下储存,随保存周数的增加,e.e.值及转化率均逐渐降低,但4周后e.e.值仍能达到90.1％,转化率保持在66.6％左右,具备较高的储存稳定性,为工业化生产(S)-乙酸苏合香酯提供了参考.     

不同植被措施下排土场边坡细沟发育时空特征

为明确不同恢复年限植被措施对煤矿排土场边坡水土流失的防治效应,采用样方调查法,以内蒙古准格尔旗永利煤矿矿区排土场边坡为研究对象,以裸露边坡(CK)为对照,研究不同恢复年限(1a、3a、5 a)的沙柳方格+沙棘+沙打旺(SHA)和沙柳方格+沙打旺(SA)2种措施对细沟发育的影响。结果表明:(1)1a时CK、SHA与SA措施细沟宽度均集中分布在4～8 cm,细沟深度均集中分布在2～4 cm;3 a时CK、SHA与SA措施细沟宽度则集中分布在4～8 cm、8～12 cm、4～8 cm,细沟深度均集中分布在4～6 cm;5 a时CK与SHA措施细沟宽度均集中分布在8～12 cm,而细沟深度则集中分布在4～6 cm和8～14 cm;(2)CK(1～5 a)、SHA措施(1～5 a)和SA措施(1～3 a)的细沟平均宽分别为7.57～11.35 cm、7.58～13.31 cm和5.57～6.14 cm,细沟平均深分别为3.38～6.23 cm、4.19～10.34 cm、2.59～4.24 cm,三者的细沟平均宽深比分别为2.39、2.12和2.05,平均细沟密度分别为1.52～5.25 m·m–2、1.42～1.68 m·m–2和1.88～2.25 m·m–2;(3)1 a时CK、SHA和SA措施的细沟宽深比随坡长变化幅度较大,随着恢复年限增加,宽深比则呈下降趋势,CK、SHA措施和SA措施的细沟密度和细沟侵蚀量均随坡长增加呈增大趋势;(4)与CK相比,1 a时SHA和SA措施边坡细沟侵蚀模数分别减小25.0%和25.86%,两种措施减蚀效果差别不大,而3 a时则分别减小了61.73%和35.31%,SHA措施减蚀效果显著增强。研究结果可以为矿区排土场边坡的植被合理布设提供科学依据与理论指导。     

EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide,Inhibits Liver Cancer Cell Growth and Adhesion

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.     

1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

文冠果优良无性系授粉组合选择及结实性状分析

  目的  探究文冠果Xanthoceras sorbifolium优良无性系最佳授粉组合，提高其坐果率、种实产量，为科学配置授粉树及优良种质资源无性系化推广提供参考。  方法  以文冠果5个优良无性系为材料，进行花期物候观测、柱头可授性及花粉活力检测。进行控制授粉，分析各授粉组合受精率、坐果率、种实产量及种实性状。  结果  ① 5个无性系开花时间在4月27日至5月14日。②雌能花开花当天及花后1 d柱头可授性较强。③雄能花初开放和完全开放当天花粉活力较高。④综合考虑各授粉组合受精率、最终坐果率、产量及种实性状得出，10号无性系较佳授粉组合为10×16、10×119，14号、15号、16号最佳授粉组合分别为14×15、15×16、16×15，10号、14号、15号、16号均不适合做119号的授粉无性系。  结论  科学的授粉树配置可以提高文冠果坐果率及种实产量。     

水霉属菌(Saprolegnia)分离培养技术研究

 对水霉(Saprolegnia)的分离培养技术进行了探索,筛选出大麻籽、油菜籽、红花籽、苍蝇、蚜虫、小麦粒、玉米片、茶叶、葵花仁9种材料作为诱发水霉菌的诱饵。其中油菜籽能防止腐霉菌(Pythium)污染,是初步纯化菌种、排除杂菌的适宜饵料。同时发现控制温度能使水霉在最短时间内分别产生无性或有性繁殖。在水培条件下,25℃左右适合菌丝及无性器官的发育,有性器官却不能产生,而只有在15~17℃条件下才能使水霉很快进入有性生殖阶段;在加入200mg/L链霉素的CMA培养基上促发有性阶段则需要在25℃左右的温度下进行,方能缩短培养时间。