The Influence for Chickens' T Lymphocyte Subsets in Peripheral Blood after Inoculated with the Recombinant Chicken Interferon-α/Chicken Interleukin-2 Complex Protein

To explore the influence of recombinant chicken interferon-α/chicken interleukin-2 fusion protein (rChIFN-α-Linker-ChIL-2 protein, recombinant fusion protein) on the percentage content of peripheral T lymphocyte subsets (PBLC) of SPF chicken,the flow cytometry was used to detected the percentage contents of CD3⁺CD4⁺, CD3⁺CD8⁺ and the CD4⁺/CD8⁺ ratio in PBLC at different days post injection in the 14-day-old SPF chicken injected with rChIFN-α-Linker-ChIL-2 protein (group 2) and recombinant chicken interferon α (rChIFN-α) protein (group 3). The results indicated that the recombinant fusion protein and rChIFN-α protein could increase the percentage content of CD3⁺CD4⁺ T lymphocyte, decrease the percentage content of CD3⁺CD8⁺ T lymphocyte, improve CD4⁺ /CD8⁺ T lymphocyte ratio during 3 to 14 d post injection. The percentage contents of CD3⁺CD4⁺, CD3⁺CD8⁺ and CD4⁺/CD8⁺ ratio in chickens of group 2 were significantly different from the PBS control group (group 1) (P<0.01), and were notably different from group 3 during 3 to 7 d post injection (P<0.05). These results showed that both the rChIFN-α-Linker-ChIL-2 protein and rChIFN-α protein could affect the percentage contents of lymphocyte subsets, improve CD4⁺/CD8⁺ ratio, enhance the cellular immune function in chicken. The influences of rChIFN-α-Linker-ChIL-2 protein on the percentage contents of CD3⁺CD4⁺ and CD3⁺CD8⁺ T lymphocyte, and CD4⁺/CD8⁺ value were notable higher than rChIFN-α, which suggested that recombinant fusion protein play a synergistic role of interferon alpha and interleukin-2 on the cellular immune function by this pathway in chicken.     

猪干扰素α-1基因的原核表达与多克隆抗体制备

[目的]利用大肠杆菌原核表达系统对猪干扰素α-1(Po IFNα-1)基因进行表达,制备Po IFNα-1重组蛋白多克隆抗体。[方法]根据Gen Bank发表的Po IFNα-1核苷酸序列,合成密码子优化的Po IFNα-1基因,构建Po IFNα-1基因的p ET-28a原核表达载体,转化大肠杆菌BL21(DE3)宿主菌后,利用IPTG进行诱导表达。表达的Po IFNα-1重组蛋白纯化后,免疫昆明系小鼠,制备多克隆抗体。采用Western blot技术检测Po IFNα-1重组蛋白的多克隆抗体与天然Po IFNα-1的反应性。[结果]合成的Po IFNα-1基因在大肠杆菌BL21(DE3)宿主菌中成功表达,针对Po IFNα-1重组蛋白的多克隆抗体能够识别天然的猪干扰素α-1。[结论]合成的Po IFNα-1基因在大肠杆菌中获得了成功表达,为后续研究奠定了基础。     

α-半乳糖苷酶基因工程菌细胞破碎条件的筛选

研究大肠杆菌表达的水稻α-半乳糖苷酶基因工程菌菌体破碎的条件。以菌液酶活性为指标,用超声波破碎、溶菌酶降解及菌体反复冻融3种方法破碎细胞。结果表明:超声波功率为200 W、破碎时间为1 min时测得的菌液酶的活性最大,为7.757 7U/m L,细胞破碎效果最好;菌体反复冻融条件下测得的菌液酶活性随冻融次数的增加而增大,但样品处理时间过长;菌液酶活性随加入溶菌酶浓度增加而增加,但有处理时间长、用量大、细胞破碎不完全的缺点。     

制备马铃薯多孔淀粉酶制剂的选择

采用α-淀粉酶和糖化酶对马铃薯淀粉进行降解制备多孔淀粉。通过对多孔淀粉的得率、吸油率的考察,研究其品质特性随不同酶用量、作用时间、温度、pH值的变化规律,并采用正交试验确定制备多孔淀粉的最佳工艺条件为:α-淀粉酶与糖化酶质量比为1∶2,且总酶量为6%,作用时间为8h,温度分别为55℃和50℃,pH值分别为6.0和3.5。     

Effects of Topographic Factors of the Site on the Essential Oil Compounds of Artemisia aucheri Aerial Parts Grown in a Mountainous Region

In this study, the chemical compositions of the essential oils from the aerial parts of Artemisia aucheri, grown in a mountainous region in Ghamsar Province, central Iran, have been analyzed by using gas chromatography (GC)–mass spectroscopy (MS) to determine how they are affected by topographic factors (site direction and elevation). Plants were sampled at random in full flowering stage in a completely randomized (CR) design with three replications. The essential oil was extracted by a modified Likens–Nickerson's simultaneous distillation–extraction (SDE) technique. Analysis of the plant oils on the sites [in two directions, north–south (N/S) and east–west (E/W)] and five elevations resulted in 30 identified compounds. Of the oil samples collected, the 99.94% of the components at the N/S site and 99.89% at the E/W site were identified. The main component in the two directions (N/S and E/W) was α-thujone with 19.5% and 18%, respectively. Other significant components in the two directions include α-pinene, davanone, camphor, and camphene. The percentages of all these main compounds on the N/S site were more than on the E/W site. Also, the main compounds increased with increasing altitude and at all five elevations. Among all the oil compounds, the percentages of α-thujone, α-pinene, and camphor were the greatest. Finally, at all the elevations, the percentage of the α-thujone was the greatest among the different oil compounds.     

四川小麦地方品种AS1643中α/β醇溶蛋白基因

用PCR方法从四川小麦地方品种AS1643中克隆到3个α/β-醇溶蛋白基因，即Gli-AS1643-1（GenBank No.DQ166376）、Gli-AS1643-2（GenBank No.DQ166377）和Gli-AS1643-3（GenBank No.DQ166378）。其中，Gli-AS1643-1和Gli-AS1643-2的编码区长度分别为873bp和852bp，可编码270和263个氨基酸残基的成熟蛋白。Gli-AS1643-3由于在编码区内有一个提前终止密码子，为不可编码成熟蛋白的假基因。序列比较显示Gli-AS1643-1、Gli-AS1643-2和 Gli-AS1643-3分别与GenBank中的α/β-醇溶蛋白基因具有较高的一致性，且序列结构非常相似。它们的N-端氨基酸序列与各种α-、β-、γ-和α/β-醇溶蛋白的基本一致，但与ω-醇溶蛋白和低分子量谷蛋白亚基的明显不同。N-端12肽串联重复紧密相关的5个脯氨酸框和类似于微卫星序列编码的2个多聚谷氨酰胺区域。在Gli-AS1643-2的N-端存在腹泻疾病活性序列，C-端含有12型腺病毒感染序列。Gli-AS1643-1、Gli-AS1643-2和Gli-AS1643-3各由6个保守的半胱氨酸残基形成3个分子内二硫键。     

棉铃虫鞣化激素基因克隆及分子特性分析

【目的】获得棉铃虫鞣化激素2个亚基(α亚基和β亚基)基因序列,分析其分子特性和表达模式,研究棉铃虫鞣化激素的作用机制奠定基础。【方法】通过生物信息学和分子生物学技术分别得到棉铃虫鞣化激素α和β亚基cDNA序列,将棉铃虫及NCBI中公布的已知其它昆虫鞣化激素α和β亚基氨基酸序列分别整理分析,利用MEGA7(7.0.14)软件的Jones-Taylor-Thornton(JTT)模型构建系统进化树并进行聚类分析,qRT-PCR分别检测两亚基在棉铃虫不同生长发育阶段的表达模式。【结果】分别获得了棉铃虫鞣化激素 α亚基与β亚基核苷酸序列。其中α亚基(GenBank登录号:AHM0247472.1)cDNA片段长694 bp,开放阅读框480 bp,编码159个氨基酸残基,预测该蛋白质分子量为17.7 kDa。该基因在卵期第3 d、1龄幼虫第1 d、3~5龄幼虫第1 d、2和3龄末蜕皮期(3M和4M)、蛹期第0 d、第3 d和第7 d表达量相对较高。β亚基(GenBank登录号:AHM0247473.1)cDNA片段长779 bp,开放阅读框420 bp,编码139个氨基酸残基,预测该蛋白质分子量为15.9 kDa。该基因在卵期至2龄末蜕皮期(3M)、3龄幼虫第2 d、4~5龄幼虫第1 d、蛹期第1 d至羽化前表达量相对较高。鞣化激素α亚基和β亚基基因均在棉铃虫胸神经节中相对表达量最高,咽下神经节次之,脑部和腹部相对表达量较弱。【结论】鞣化激素在鳞翅目昆虫中具有较高的保守性;α亚基和β亚基基因主要在棉铃虫卵期、初孵幼虫期、蛹期和新羽化成虫期发挥作用;鞣化激素在棉铃虫幼虫体内主要由胸部神经节转录合成。     

α预不变凸函数的若干性质

考虑了一类广义凸函数———α预不变凸函数,利用中间点的α预不变凸性,讨论了α预不变凸函数与半连续函数之间的关系,得到了α预不变凸函数的一些性质。     

Comparison of serum lipid compositions, lipid peroxide, alpha-tocopherol and lipoproteins in captive marine mammals (bottlenose dolphins, spotted seals and West Indian manatees) and terrestrial mammals

Concentrations of serum lipid components, lipid peroxide (LPO) and α-tocopherol and electrophoretic patterns of lipoproteins in serum samples obtained from captive marine mammals and terrestrial mammals were compared. Serum concentrations of total cholesterol, free fatty acid, and phospholipid in fish-eating animals were significantly higher than those in manatees and cows. Serum LPO and α-tocopherol concentrations in the fish-eating animals were also significantly higher than those in manatees, cows and dogs. Different patterns of densitometric scans of low density lipoprotein (LDL) and a significantly lower percentage of LDL were demonstrated in the dolphins compared with the seals, cow and dogs. The concentration of LPO was significantly correlated with triglyceride and phospholipid concentrations in serum from the dolphins. These results suggest that triglyceride and phospholipid are susceptible to oxidative reaction in fish-eating animals. Evaluation of serum lipids, LPO and α-tocopherol concentrations is needed for nutritional husbandry for fish-eating animals.     

Absorption of Different Oral Formulations of Natural Vitamin E in Horses

The objective of this study was to determine the effect of vitamin E source on plasma vitamin E concentration. Five different formulations of natural source vitamin E (4,000 IU/day) were supplemented to Standardbred mares (n = 4 per group) for 14 days. Treatment 1 was given 10 g of 400 IU/g natural acetate (RRR-α-tocopheryl-acetate) powder, treatment 2 was given 6.66 g of 600 IU/g natural acetate powder, treatment 3 was given 6.66 g of 600 IU/g natural alcohol (RRR-α-tocopherol) powder, treatment 4 was given 20 g of 200 IU/g micellized natural alcohol powder, and treatment 5 was given 8 mL of 500 IU/mL micellized natural alcohol liquid. Blood samples were collected before supplementation (day 0), after day 7 and day 14 of supplementation, and analyzed for plasma α-tocopherol. Treatments 1, 2, and 3 increased (P < .05) from days 0 to 7, but remained similar at day 14. Treatments 4 and 5 also increased (P = .004, and P < .0001, respectively) from days 0 to 7 and were higher (P < .05) than treatment 1, 2, and 3. Plasma levels peaked at day 7 and either plateaued or decreased (treatment 5, P = .004) at day 14. The micellized alcohol formulations remained elevated (treatment 4) or decreased (treatment 5) after achieving peak blood concentration and, as hypothesized, tended to have plasma concentrations higher than the other vitamin E forms. Care should be taken to ensure that the most active biologically available form of vitamin E is provided in the diet to maximize its efficiency.