花生壳黄酮的提取纯化工艺

本文采用酶法预处理结合超声波辅助提取的方式，最大程度地提高了花生壳总黄酮的得率，并优化大孔树脂纯化工艺，提高了有效成分的纯度。花生壳黄酮的最佳提取工艺为：花生壳粉与水混合，半纤维素酶与木聚糖酶按1:1（m/m）复配，用量0.25‰，50℃酶解30 min后，按料液比1:20（m/V）加入乙醇至终浓度60%，于功率1000 W，55℃超声波辅助提取60 min，花生壳总黄酮的得率约为2.5%。选用D101型大孔树脂，上样缓冲液为pH 5.0的60%乙醇溶液，洗脱液为pH 10.0的70% 乙醇溶液，上样与洗脱流速为0.75 BV/h和1.5 BV/h。纯化后的花生壳总黄酮和木犀草素的纯度分别为10.54%和5.85%，提高了90%和120%。     

Pharmacokinetics and Pharmacodynamics of a Depolymerized Glycosaminoglycan from Holothuria fuscopunctata,a Novel Anticoagulant Candidate,in Rats by Bioanalytical Methods

dHG-5 (Mw 5.3 kD) is a depolymerized glycosaminoglycan from sea cucumber Holothuria fuscopunctata. As a selective inhibitor of intrinsic Xase (iXase), preclinical study showed it was a promising anticoagulant candidate without obvious bleeding risk. In this work, two bioanalytical methods based on the anti-iXase and activated partial thromboplastin time (APTT) prolongation activities were established and validated to determine dHG-5 concentrations in plasma and urine samples. After single subcutaneous administration of dHG-5 at 5, 9, and 16.2 mg/kg to rats, the time to peak concentration (T_max) was at about 1 h, and the peak concentration (C_max) was 2.70, 6.50, and 10.11 μg/mL, respectively. The plasma elimination half-life(T_1/2β) was also about 1 h and dHG-5 could be almost completely absorbed after s.c. administration. Additionally, the pharmacodynamics of dHG-5 was positively correlated with its pharmacokinetics, as determined by rat plasma APTT and anti-iXase method, respectively. dHG-5 was mainly excreted by urine as the unchanged parent drug and about 60% was excreted within 48 h. The results suggested that dHG-5 could be almost completely absorbed after subcutaneous injection and the pharmacokinetics of dHG-5 are predictable. Studying pharmacokinetics of dHG-5 could provide valuable information for future clinical studies.     

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.     

玉米秸秆基纤维素保水剂对土壤持水性能及冬小麦根系生长的影响

为探讨自制的玉米秸秆基纤维素保水剂(SAP)在农业生产上的应用潜力,利用扫描电镜表征了SAP吸水前后形貌变化,探究了SAP施用量对土壤持水性能和不同程度水分胁迫下冬小麦根系生长的影响。结果表明：SAP在土壤中吸水膨胀后降低了土壤容重,土壤最大水分含量较不施SAP显著提高19.1%~67.6%,同时降低了前3天土壤水分蒸发速率。轻度（60%~70%田间持水量）和中度（40%~50%田间持水量）水分胁迫下,施用0.4% w/w SAP处理冬小麦总根长提高15.9%和24.7%,根系活力提高29.6%和43.5%,但充足供水（75%~90%田间持水量）时施用SAP对冬小麦根系活力无显著影响。因此,玉米秸秆基纤维素保水剂施入土壤后能够改善土壤持水性能,从而缓解水分胁迫对冬小麦根系生长的抑制效应。     

园林绿化废弃物覆盖对城市绿地土壤肥力影响

  目的  本研究旨在探讨园林绿化废弃物覆盖对城市绿地土壤肥力影响，以期为园林绿化废弃物资源化利用解决城市绿地土壤问题提供依据。  方法  本研究以北京市典型城市绿地土壤?圆明园为研究对象，通过野外试验设置4种不同处理:不覆盖（CK）、上层木片 + 下层堆置15 d的园林绿化废弃物堆肥（处理A）、上层木片 + 下层堆置30 d的园林绿化废弃物堆肥（处理B）、上层木片 + 下层堆置60 d的园林绿化废弃物堆肥（处理C），进行室内分析测定，结合修正的内梅罗指数法计算土壤综合肥力指数，运用Pearson相关性及逐步回归分析，分析不同覆盖处理对土壤综合肥力的影响。  结果  0 ~ 10 cm土层中，C覆盖处理显著提高土壤有效磷、速效钾含量（P < 0.05）；10 ~ 20 cm土层中，C覆盖处理显著提高土壤速效钾含量（P < 0.05）。土壤综合肥力指数显示，C覆盖处理显著提高0 ~ 10 cm土层的综合肥力（P < 0.05）。修正的内梅罗指数法中F_imin值及逐步回归分析表明，土壤综合肥力限制因子为土壤密度及有效磷，土壤综合肥力最主要影响因子为有机质，其次为土壤有效磷及土壤密度。  结论  综上，园林绿化废弃物覆盖对土壤理化性质及综合肥力有不同程度的影响，结果显示处理C为最佳覆盖方案，今后研究中要着重注意土壤肥力主要影响因子（土壤有机质、土壤密度及有效磷）的动态变化及改善。      

斜纹夜蛾Toll受体基因的全基因组分析及其在杀虫剂抗性中的作用

利用公布的斜纹夜蛾基因组对TLRs基因进行全基因组分析,应用生物信息学工具分析斜纹夜蛾TLRs家族基因成员在鳞翅目害虫TLRs基因家族中的进化关系、组织表达特异性,结合转录组数据对农药反应敏感的TLRs进行基因表达及其调控网络进行分析,分析斜纹夜蛾Toll受体基因在杀虫剂抗性中的作用.结果表明:共分析出了7个TLRs基因,斜纹夜的TLRs家族基因与棉铃虫TLRs家族基因同源性较高.7个基因中的1个基因SLTLR3在肠道中高度表达,有5个TLRs基因对农药处理反应敏感.对农药反应敏感的TLRs进行基因表达和其网络进行分析,只发现了1个TLRs(SLTLR7)和其他30个基因之间的存在共表达关系,这些基因主要涉细胞表面受体信号途径、细胞定位、信号传导、细胞调控过程、生物调控过程.     

Molecular Characterization of the Von Willebrand Factor Type D Domain of Vitellogenin from Takifugu flavidus

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val¹¹⁵, ASP¹¹⁶, Val¹¹⁷, and Lys¹²²) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein–toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.     

利用HPLC-ICP-MS方法测定中华绒螯蟹不同形态砷的残留量

采用高效液相色谱串联电感耦合等离子体质谱仪(HPLC-ICP-MS)测定中华绒螯蟹可食部位6种砷形态的残留量。经过前处理、提取和脱脂后,通过色谱柱、流动相、pH和流速等参数调节,筛选得到最优化的分离和检测条件。结果表明,6种砷形态的混合标准溶液在质量浓度0~50 μg/L范围时,均呈现良好的线性关系,其R²达到0.999,检测限为0.0060~0.0912 μg/kg,定量限为0.020~0.304 μg/kg。在样品中添加0.5、5、50 μg/L浓度水平,回收率良好,范围是70%~120%,相对标准偏差(RSD)均<5%,方法精密度与重复性均良好。本方法为中华绒螯蟹可食部位砷形态残留量建立了有效检测方法,为水产品砷残留快速检测提供技术支撑。     

Multiplication of the Recombinant Strain Re-7 of Avian Influenza Virus Subtype H5 in MDCK Cells

This study was conducted to explore the multiplication pattern of the recombinant strain Re-7 of avian influenza virus subtype H5 in Madin Darby Canine Kidney(MDCK) cells and to determine the optimal multiplicity of infection(MOI) and the optimal time for virus harvest. The recombinant strain Re-7 was inoculated at different MOIs into MDCK cells grown in serum-free medium in 100 L bioreactors for replication. Then,the hemagglutination(HA) titer, 50% tissue culture infectious dose(TCID50) and 50% embryo infectious dose(EID50) of culture medium were measured once every 12 h from 24 h after virus inoculation to determine the optimal MOI. After that, virus was inoculated at the optimal MOI deter-mined above into MDCK cells for large-scale virus replication to determine the optimal time for virus harvest. The results showed that the optimal MOI was 10~(-2), and the optimal time for virus harvest was 60 h after inoculation. Under these conditions, the HA titer, TCID_(50) per 1 m L and EID50 per 0.1 m L were increased to 1∶102 4, 10~(7.33) and 10~(6.83), respectively. This study provides relatively stable parameters for large-scale production of the recombinant strain Re-7 of avian influenza virus subtype H5.     

Optimizing the Proportions of Plant Protein Ingredients in the Diet of Juvenile Blunt Snout Bream,Megalobrama amblycephala

An 8‐wk feeding trial was conducted to evaluate the influence of different proportions of dietary plant ingredients on the growth of juvenile blunt snout bream, Megalobrama amblycephala. A 2 × 3 factorial arrangement was set for this trial with three ratios of cottonseed meal (CSM) and canola meal (CM) (2:1, 1:1, or 1:2) and three proportions of soybean meal (SBM) (2, 1, or 0). The results showed that the specific growth rate and protein efficiency ratio both significantly decreased with decreasing amounts of SBM in the diets of blunt snout bream (P < 0.05), while the feed efficiency ratio and feed intake increased significantly (P < 0.05). Meanwhile, the growth performance and feed utilization were shown to improve with increasing amounts of CSM, although the effects were not significant (P > 0.05). The interaction of these two factors had no effect on either growth or feed utilization (P > 0.05). Diet composition was not seen to affect body composition (P > 0.05). The ratio of CSM to CM significantly affected gut amylase activity, liver alanine aminotransferase content, and plasma cholesterol levels (P < 0.05). The interaction between the two main effects significantly influenced the plasma glucose level (P < 0.05). Therefore, the suitable proportion of plant feed for blunt snout bream should include high levels of SBM and CSM, with a SBM : CSM : CM ratio of 2:2:1.