橘色双冠丽鱼黑素皮质素受体1基因(MC1R)整胚原位杂交

为探究黑素皮质素受体1基因(MC1R)在橘色双冠丽鱼胚胎发育和体色形成过程中的表达定位及功能,本研究首先制备了MC1R基因的RNA正反义探针,T7方向转录的正义RNA探针质量浓度为447.529 ng/μL,SP6方向转录的反义RNA探针质量浓度为342.698 ng/μL。经10～20倍稀释后的探针用于原位杂交,MC1R基因探针表达定位显示,随橘色双冠丽鱼胚胎的发育杂交信号总体呈逐渐减弱趋势,原肠期和视泡期其卵黄和胚体侧卧部分有杂交信号分布;出膜期其脊柱、卵黄囊及其内容物和色素细胞内出现杂交信号;出膜期第1天,卵黄表面有信号分布。总体来看,杂交信号在脊索、卵黄囊、卵黄囊内容物质及色素细胞有特异表达,而以正义探针作为阴性对照组在5个胚胎阶段均无任何信号,杂交信号显现的MC1R表达定位说明其在橘色双冠丽鱼色素细胞的分化、迁移中起到重要调控作用,也与神经、营养、免疫功能相关,初步建立了鱼类体色相关基因功能和定位研究的整胚原位杂交方法。     

水温对怀头鲇血液学指标的影响

本实验设定4℃、9℃、14℃、18℃、24℃和30℃6个水温梯度,测定了水温变化对二龄怀头鲇血液生理生化指标的影响。结果显示,水温4～30℃范围内,怀头鲇血红蛋白含量随着水温的升高呈波浪型变化,但变化幅度较小;而血沉与血红蛋白含量变化趋势相反,变化幅度较大;血液中的红细胞数量随水温的升高先增加再减少;血清中的总蛋白、白蛋白和球蛋白的变化趋势一致,都是随水温的升高其含量先增加再减少,而且在9～18℃区间变化较急剧;血清中的血糖,高密度脂蛋白,谷草转氨酶,三碘甲状腺氨酸,甲状腺素,促甲状腺激素,钾和镁八项指标在各水温之间不存在显著差异（P〉0．05）;血清中的尿素氮,甘油三脂,总胆固醇,低密度脂蛋白,谷丙转氨酶,碱性磷酸酶,钠,钙,氯化物和渗透压在部分水温之间存在显著差异（P〈0．05）。     

Changes in the Quality of Refined Fish Oil in an Accelerated Storage Study

ABSTRACT

The aim of this study was to prepare the refined oil derived from Sardinella longiceps by a step-wise procedure of degumming, bleaching, and deodorization. The refined sardine oil showed greater susceptibility toward oxidation than the crude oil, but the addition of ethyl acetate fractions of the red seaweeds Hypnea musciformis, Kappaphycus alvarezii, and Jania rubens with great antioxidant potential significantly increased the oxidative stability of the refined oil. The refined oil supplemented with the ethyl acetate fraction of these seaweeds indicated more noteworthy oxidative stability indices (≥ 0. 51 h) than α-tocopherol. The induction time was greater for refined oil added with H. musciformis (1.26 h) than butylated hydroxytoluene (1.04 h) and α-tocopherol (0.4 h). No traces of aromatic (δ 6.7–7.5 ppm), hydroperoxides, and aldehydes (δ 9–10 ppm) in the ¹H-NMR spectra of the refined oil supplemented with seaweed extracts at the baseline and after the accelerated storage study demonstrated that the seaweed extracts were able to prevent the formation of secondary oxidation products in the unsaturated system during storage. This study demonstrated the potential of seaweeds as natural alternatives to the synthetic antioxidants to prevent the rancidity of the refined fish for use in food and pharmaceutical industries.     

Effect of dietary protein:energy ratio on intake, growth and metabolism of juvenile green abalone Haliotis fulgens

Juvenile green abalone Haliotis rufescens were grown under laboratory conditions at 21±1 °C and fed formulated diets consisting of different protein:energy ratios (mg protein/kcal), 62, 74, 85, 100, 108, for 60 days. The level of crude protein ranged from approximately 26% to 44% while the energy content remained constant at about 4.1 kcal g⁻¹. Growth ranged from 3.63 to 12.33 mg day⁻¹. The growth of abalone fed the 100 and 108 diets was significantly greater than that of each of the other diets. Protein efficiency ratio increased as the dietary protein content increased except for the T108 diet (44% crude protein). Abalone apparently consume food to satisfy an energy requirement. Caloric expenditure due to metabolism was estimated for abalone fed diets with protein ratios of 62, 85, 100. Energy loss due to respiration did not vary appreciably among abalone fed the different diets. The proportional distribution of dietary energy into fecal, digestible, growth, and metabolic energy was estimated for abalone fed these diets. Apparent dry matter digestibility was among the lowest for abalone fed the 100 P:E diet, but growth of abalone fed this diet was significantly higher than that of each of the other treatments except the 108 diet. Unexplained energy loss to achieve balance ranged from 7% to 28.5%, some of which is probably due to differential mucus and ammonia production. Results suggest a potential for the reduction of both dietary protein and lipid without causing any adverse effects on the growth response.     

野生和“黄海1号”中国明对虾不同组织基因组DNA的MSAP分析

为了从表观遗传学角度讨论中国明对虾(Fenneropenaeus chinensis)野生群体和人工选育新品种“黄海1号”不同组织间甲基化水平和多态性差异,应用甲基化敏感扩增多态性(mehylation sensitive amplified polymorphism, MSAP)分别对野生群体组中国明对虾和“黄海1号”肌肉、鳃、血液3种组织样品基因组DNA的CCGG甲基化水平进行对比分析,试图从表观遗传学角度探讨影响中国明对虾生长性状的分子机制.采用30对引物进行选择性扩增,经聚丙烯酰胺凝胶电泳(Polyacrylamide Gel Electrophoresis, PAGE)带型结果显示,野生群体组中国明对虾肌肉、鳃和血液的甲基化比例分别为23.1%,22.3%和19.7%;而“黄海1号”肌肉、鳃和血液的甲基化比例分别为21.4%、19.6%和18.9%.野生群体组中国明对虾和“黄海1号”同一组织间的甲基化水平和甲基化多态性水平不同,肌肉、鳃和血液不同组织间的甲基化水平和甲基化多态性水平亦不同.DNA 甲基化多态性带型分析显示,鳃组织的甲基化水平和多态性水平在野生群体组中国明对虾和“黄海1号”间变化趋势最大,肌肉最稳定.本研究旨为甲基化修饰与中国明对虾生长性状间的相关性研究提供依据.     

Hmgn3和Hoxa10对小鼠子宫基质细胞蜕膜化过程中Foxo1表达的调控

利用小鼠子宫基质细胞体外诱导蜕膜化模型转染Hmgn3或Hoxa10过表达载体及siRNA片段,通过荧光定量PCR方法检测Hmgn3和Hoxa10对Foxo1表达的调控。结果显示:转染Hmgn3或Hoxa10过表达载体可促进Foxo1在子宫基质细胞蜕膜化过程中的表达,而转染Hmgn3或Hoxa10siRNA则可抑制Foxo1的表达。在小鼠子宫基质细胞中转染Hmgn3或Hoxa10siRNA后再添加孕酮,Foxo1的表达显著下降。同样,干扰Hmgn3或Hoxa10也可减低cAMP对Foxo1表达的调控。结果表明:Hmgn3和Hoxa10可通过Foxo1来影响小鼠子宫基质细胞的蜕膜化,孕酮和cAMP可通过Hmgn3和Hoxa10来调控Foxo1在子宫基质细胞中的表达。     

Atorvastatin inhibits IL-1β and IL-18 releases via lowering the expression of NLRP1 inflammasome

AIM: To explore the role of nucleotide-binding oligomerization domain-like receptor protein 1 (NLRP1) inflammasome in atorvastatin-induced reduction of interleukin-1β (IL-1β) and interleukin-18 (IL-18) releases from the THP-1 macrophages. METHODS: Lipopolysaccharide (LPS, 10 μg/L) was used to trigger the secretion of IL-1β and IL-18 in the THP-1 macrophages. The cells were incubated with different concentrations of atorvastatin (1, 10 and 20 μmol/L) for 24 h, or treated with 10 μmol/L atorvastatin for different time (12 h, 24 h and 48 h). NLRP1 siRNA was transfected into the THP-1 cells. The mRNA expression of NLRP1 inflammasome was detected by RT-PCR. The protein expression of NLRP1 inflammasome was determined by Western blot. The secretion of proinflammatory cytokines IL-1β and IL-18 was quantified by ELISA. RESULTS: Atorvastatin inhibited the mRNA and protein expression of NLRP1 inflammasome in the THP-1 macrophages in a dose- and time-dependent manner. Transfection of NLRP1 siRNA significantly decreased the protein expression of NLRP1 and promoted the suppressive effect of atorvastatin on IL-1β and IL-18 secretion in the THP-1 macrophages. CONCLUSION: Atorvastatin inhibits the production of IL-1β and IL-18 in the macrophages through decreasing NLRP1 inflammasome expression, possibly contributing to the anti-inflammatory effect of atorvastatin on atherosclerosis.     

Functional changes of retinal ganglion cells in rd1 mice during midterm of retinal pigmentosa

AIM: To investigate how the function of retinal ganglion cells (RGCs) change in the midterm of retinal pigmentosa (RP) in rd1 mice (a transgenic animal model of RP). METHODS: The action potentials from multiple RGCs in rd1 mice at postnatal 20 d (P20) or normal C57 mice (control) were simultaneously recorded by multi-electrode array recording. The functional changes of surviving ganglion cells were evaluated by comparing spontaneous and light-evoked activities of RGCs between rd1 and control mice. The extent of photoreceptor degeneration was verified by immunohistochemical staining. RESULTS: Immunohistochemistry results showed the thickness of the retinal photoreceptor layer of rd1 mice was significantly lower than that in normal mice at P20. According to the light response properties, we classified ganglion cells into 6 subgroups: ON sustained, ON transient, ON-OFF sustained, ON-OFF transient, OFF sustained and OFF transient RGCs, with a very tiny percentage of OFF sustained RGCs (1.0%~3.1%). The percentage of RGCs remaining light responsive in rd1 mice was significantly lower than that in C57 mice. The average spontaneous spiking rate for rd1 RGCs was overall significantly increased compared to that in C57 cells, whereas different RGC types had different changes. The light-induced responses and light sensitivities of all types of RGCs in rd1 mice were both significantly lower than those in C57 mice. CONCLUSION: The photoreceptors of rd1 mice are severely degenerated in the midterm of retinal degeneration. The functions of RGCs in rd1 mice in the midterm of degeneration decay obviously, with variance in different RGCs types.     

“紫科1号”红豆杉微枝繁殖技术研究

以"紫科1号"红豆杉的6种半木质化枝条为扦插繁殖材料,经3种处理液预处理,在8种基质上进行扦插繁殖试验。结果表明:3~12 cm长的枝条均可用作插穗,选用长5 cm左右的枝条进行微枝繁殖较为节约材料;用IBA∶NAA=3∶2,总溶质浓度为1 000 mg/L的处理液速蘸基部后,插于黄心土∶河沙=3∶2的基质中,其后每周用稀释1 000倍的处理液喷洒1次,生根率平均可达97.36%。     

动物鲜味受体的研究进展及其基因表达调控

动物的鲜味受体包括代谢型谷氨酸受体(m GluR)和味觉受体异源二聚体(T1R1/T1R3),是C型G蛋白偶联受体,N末端捕蝇草模块(VFT)区域可与鲜味配体结合,识别鲜味。本文主要论述了鲜味受体的研究进展、鲜味识别转导机制及鲜味受体基因的表达调控等,以期为相关研究提供参考。