Construction and characterization of an HCV-derived multi-epitope peptide antigen containing B-cell HVR1 mimotopes and T-cell conserved epitopes

Hepatitis C (HCV) genome is highly variable, particularly in the hypervariable region 1 (HVR1) of its E2 envelope gene. The variability of HCV genome has been a major obstacle for developing HCV vaccines. Due to B-cell HVR1 mimotopes mimicking the antigenicity of natural HVR1 epitopes and some T-cell epitopes from the consensus sequence of HCV genes conserving among the different HCV genotypes, we synthesized an minigene of HCV-derived multi-epitope peptide antigen (CMEP), which contains 9 B-cell HVR1 mimotopes in E2, 2 conserved CTL epitopes in C, 1 conserved CTL epitope in NS3 and 1 conserved Th epitope in NS3. This minigene was cloned into a GST expression vector to generate a fusion protein GST-CMEP. The immunogenic properties of CEMP were characterized by HCV infected patients’ sera, and found that the reactivity frequency reached 75%. The cross reactivity of anti-CEMP antibody with different natural HVR1 variants was up to 90%. Meanwhile, we constructed an HCV DNA vaccine candidate, plasmid pVAX1.0-st-CMEP carrying the recombinant gene (st) of a secretion signal peptide and PADRE universal Th cell epitope sequence in front of the CMEP minigene. Immunization of rabbits with pVAX1.0-st-CMEP resulted in the production of antibody, which was of the same cross reactivity as the fusion protein GST-CMEP. Our findings indicate that the HCV-derived multi-epitope peptide antigen in some degree possessed the characteristics of neutralizing HCV epitopes, and would be of the value as a candidate for the development of HCV vaccines.     

荣成天鹅湖湿地越冬大天鹅食性分析

2005年12月～2006年3月以大天鹅(Cygnus cygnus)为对象,研究其越冬期间的食性,利用粪便显微组织分析法定量调查其食物组成。结果表明,大天鹅的越冬食物主要为小麦(Triticum aestivum)、海带(Laminaria japonica)和大叶藻(Zostera marina),其中,小麦的比例最大,分别占冬季食物组成的99.67%和早春的90.17%。对冬季与早春的食物组成的差异进行分析,发现大天鹅在天鹅湖保护区发生了食物替代现象。结合以往资料,认为生态污染和湿地结构性破坏导致大天鹅传统食物减少,致使食物替代现象的发生。     

不同微生物菌剂对基质酶活性和番茄产量及品质的影响

以鸡粪配方基质(腐熟鸡粪:腐熟玉米秸秆:河沙体积为3:4:3)和牛粪配方基质(腐熟牛粪:腐熟玉米秸秆:河沙体积为3:4:3)为试材,研究了基质中分别添加地福来、酵素菌、EM菌、枯草芽孢杆菌和农用微生物菌剂对基质酶活性和番茄产量及品质的影响.结果表明: 两种配方基质添加微生物菌剂40和60 d时根际基质的脲酶、蔗糖酶和碱性磷酸酶活性均显著提高,番茄植株生长量、果实产量及维生素C含量显著高于对照.鸡粪和牛粪配方基质均以添加地福来效果最好,番茄单株产量分别较各自对照增加14.7%和40.0%,果实维生素C含量分别提高22.2%和39.7%.在不添加微生物菌剂的情况下,鸡粪配方基质栽培的番茄单株产量和果实维生素C含量高于牛粪配方基质;分别添加地福来后,两种基质栽培的番茄单株产量和果实维生素C含量无显著差异.     

大仓鼠的核型与B染色体研究

采用骨髓细胞染色体制片法,对分布于山东济南、泰山、东北长白山和陕西西安的大仓鼠的染色体组型、Ｇ－带、Ｃ－带和银染核型进行了分析研究。济南、西安和长白山的标本的二倍体数目和核型相似,２ｎ＝２８,２２ｔ＋４ｍ＋ＸＹ（ｓｔ,ｍ）。泰山标本的二倍体数目为２ｎ＝２８～２９,即在６７５％的中期相中多出了一条形态最小的端着丝粒染色体,这条染色体为Ｂ染色体,可能起源于Ｘ染色体。泰山标本的Ａ染色体组与上述３地标本相同。４地标本的Ｇ－带、Ｃ－带和银染核型相似。除Ｂ染色体外,每个端着丝粒染色体都具有着丝粒异染色质,ＡｇＮＯＲｓ较恒定地出现在Ｎｏｓ２,４,８,９,１３染色体上。也就是说大仓鼠的Ｂ染色体为Ｃ－带阴性,不携带核仁组织者。这种Ｂ染色体Ｃ－带阴性的特征在赤狐、黑家鼠和大林姬鼠朝鲜亚种中亦有报道。     

γ-干扰素时间分辨免疫荧光分析方法的建立

采用生物素-Eu³⁺标记链亲和素双抗体夹心时间分辨免疫荧光分析技术(TRIFMA),建立新的γ-干扰素检测技术,提高γ-干扰素检测方法的灵敏度.用固相包被的兔多抗捕获样品中IFN-γ,以具有中和IFN-γ抗病毒活性的生物素化单抗作为二抗体,再加Eu³⁺标记链亲和素并荧光检测.已知不同浓度标准IFN-γ CPS值的标准曲线,判断待检样品中IFN-γ量.本方法最低检测值为0.02 μg/L,检测范围为0.02～400 μg/L,而TNF-α,IL-2和IFN-α等细胞因子无交叉反应.对基因工程IFN-γ的生产,纯化过程中定性, 定量监控以及对培养细胞上清中IFN-γ量的检测等都有实用价值.     

逆转录病毒载体介导胸苷磷酸化酶在胰腺癌细胞表达

人胸腺嘧啶核苷磷酸化酶（ＴＰ）在一些肿瘤组织中活性增高,但其功能目前了解尚少。构建了表达ＴＰ的重组逆转录病毒载体,直接导入人胰腺癌ＰＣ－２细胞,ｍＰＣＲ扩增、Ｓｏｕｔｈｅｒｎ及Ｎｏｒｔｈｅｒｎ印迹和原位杂交证实转染细胞有外源ＴＰ的整合及表达,酶活性检测发现含外源ＴＰ细胞ＴＰ活性比ＰＣ－２细胞的内源性ＴＰ活性高ＴＰ活性高４０－７０倍,生长曲线和＾３Ｈ－ＴｄＲ参入率检测未发现含外源ＴＰ细胞生物学行为的     

Effects of Calcium on Absorption and Distribution of Ions and H+-ATPase Activity in Barley and Wheat Under Salt Stress

Calcium decreased Na+ absorption and transportation to the shoots,increased K+ and Ca2+ absorption and transportation ,decreased the leakage of electrolyties,and increased the accumulation of dry matter in barley and wheat seedlings under NaC1 stress. Calcium ion promoted the H+-ATPase activities in plasma membrane and tonoplast vesicles isolated from the young roots of the two plants, and increased respiration of the roots. This is in consistent with the results that calcium regulates ion absorption and distribution via its enhancement of H+-ATPase activities in plasma membrane and tonoplast.     

Pollination Biology in Cimicifuga nanchuanensis,an Endangered Species (Ranunculaceae)

The pollination biology of Cimicifuga nanchuanensis Hsiso, an endangered species and C. siraplex Wormsk, a widespread species as a control, has been studied. Protandry occurs in both species, the former goes further than the latter. The breeding system of C. nanchuanensis is dominantly xenogamous. The average visitation rate of flies as the main pollinators is 1.1 and 0.6 per flower per day at anthesis and after stamen shedding respectively. C. simplex is mainly an entomophilous plant. However, wind pollination could occur within a short distance. Moreover, a certain ratio of self pollination exists also in this species. The mean visitation rates of flies and bees, the main pollinators of C. simplex, are 1.4 and 1.2 respectively at anthesis. The pollinating insects of both plant species are attracted by their big inflorescences and the fragrant odor. Nectar, the reward for pollinators, produces in the secretory cells at the base of the spoonform staminode. In natural conditions, an average of 17 pollen grains are found on the surface of a stigma resulting in a mean seed set rate of 45.9 %. Pollination limitation (very few pollinators and their low efficiency of pollination) in C. nanchuanensis is one of the weak links in its life cycle and one of the reasons for the endangerment of this species.