西瓜细菌性果斑病菌基因Aave-3192和Aave-2108的功能研究

从本实验室构建的西瓜细菌性果斑病菌菌株A13的Mini-Tn5转座子突变体库中筛选出2株致病性丧失的突变菌株166和167,亚克隆及测序结果表明,突变株166和167的Tn5插入位点基因编号分别为Aave-3192和Aave-2108。为了进一步明确这2个基因的功能,分别对这2个基因进行互补。生物学试验结果表明,互补菌株恢复对西瓜果实的致病性,且在游动性、群体感应、胞外多糖、生物膜等性状上部分恢复至野生菌株A13的水平,表明这2个基因与该病菌在西瓜上的致病性相关。     

芩连液与白虎汤对气分证家兔补体经典途径活化的影响比较

为比较经方白虎汤与自拟方芩连液的分子疗效区别,以静脉注射内毒素建立了具有高热证候的气分证家兔模型,并用白虎汤和自拟方芩连液分别进行治疗,通过ELISA检测比较治疗前后血清总补体(CH50)的含量变化,并采用实时荧光定量PCR技术检测了肝组织中补体第三成分(C3)基因和C-反应蛋白(CRP)基因的表达变化。结果表明,在气分证家兔模型中,补体系统以经典途径活化。白虎汤的治疗能同时保证C3基因和CRP基因的高表达起到治愈作用,而芩连液仅以提高CRP的表达量发挥一定的保护作用。研究表明白虎汤对动物气分证的疗效优于芩连液。     

Dysferlin and animal models for dysferlinopathy

Dysferlin (DYSF) is involved in the membrane-repair process, in the intracellular vesicle system and in T-tubule development in skeletal muscle. It interacts with mitsugumin 53, annexins, caveolin-3, AHNAK, affixin, S100A10, calpain-3, tubulin and dihydropyridine receptor. Limb-girdle muscular dystrophy 2B (LGMD2B) and Miyoshi myopathy (MM) are muscular dystrophies associated with recessively inherited mutations in the DYSF gene. The diseases are characterized by weakness and muscle atrophy that progress slowly and symmetrically in the proximal muscles of the limb girdles. LGMD2B and MM, which are collectively termed "dysferlinopathy", both lead to abnormalities in vesicle traffic and membrane repair at the plasma membrane in muscle fibers. SJL/J (SJL) and A/J mice are naturally occurring animal models for dysferlinopathy. Since there has been no an approach to therapy for dysferlinopathy, the immediate development of a therapeutic method for this genetic disorder is desirable. The murine models are useful in verification experiments for new therapies and they are valuable tools for identifying factors that accelerate dystrophic changes in skeletal muscle. It could be possible that the genetic or immunological background in SJL or A/J mice could modify muscle damage in experiments involving these models, because SJL and A/J mice show differences in the progress and prevalent sites of skeletal muscle lesions as well as in the gene-expression profiles of their skeletal muscle. In this review, we provide up-to-date information on the function of dysferlin, the development of possible therapies for muscle dystrophies (including dysferlinopathy) and the detection of new therapeutic targets for dysferlinopathy by means of experiments using animal models for dysferlinopathy.     

The effect of time‐dependent protein restriction on growth factors,nonspecific immunity,body composition,fatty acids and amino acids in the Siberian sturgeon (Acipenser baerii)

The effect of time‐dependent protein restriction (PR) and refeeding were investigated on growth, body composition, fatty acids (FA), amino acids (AA) and nonspecific immune functions in the juvenile Siberian sturgeon (Acipenser baeri; 75 ± 5 g). Test diets included: T1 (control) and T2 fish fed diets containing 400 and 300 g/kg protein respectively during the whole experimental period, T3 (every other day) and T4 (every other week) fish were fed diets containing 300 g/kg protein (PR) and refeeding with diet containing 400 g/kg protein respectively, and T5 fish fed a diet containing 300 g/kg protein for 3 weeks and a diet containing 400 g/kg protein for 5 weeks, were fed to the fish for 56 days by visual satiation. Unlike ration T2 and T5, feeding treatments of T3 and T4 showed an increase in fish growth and body composition (p < 0.05) and they were very close to the control group (p > 0.05). Regarding the fatty acid profile, although the percentage of highly unsaturated fatty acids (HUFA) and polyunsaturated fatty acids (PUFA) declined significantly in the T2 and T5 groups, the saturated fatty acid (SFA) and monounsaturated fatty acids (MUFA) indicated no change among the treatments. In addition, the T2 and T5 groups demonstrated a reduction in essential amino acids (EAA) with a simultaneous increase in nonessential amino acids (NEAAs), which was significant from that of controls and other groups (p < 0.05). In terms of nonspecific immune parameters (serum lysozyme activity and alternate complement activity [ACH50]), treatment 3 has an appropriate result that did not have a significant difference with the control group (p > 0.05), but there was significant difference with other groups (p < 0.05). As a result, the T3 treatment can be used in sturgeon aquaculture practically without any negative impacts on biological or physiological indices.     

Multi-variable grey model (MGM (1,n,q)) based on genetic algorithm and its application in urban water consumption

Urban water consumption has some characteristics of grey because it is influenced by economy, population, standard of living and so on. The multi-variable grey model （MGM（1,n））, as the expansion and complement of GM（1,1） model, reveals the relationship between restriction and stimulation among variables, and the genetic algorithm has the whole optimal and parallel characteristics. In this paper, the parameter q of MGM（1,n） model was optimized, and a multi-variable grey model （MGM（1 ,n,q）） was built by using the genetic algorithm. The model was validated by examining the urban water consumption from 1990 to 2003 in Dalian City. The result indicated that the multi-variable grey model （MGM（1,n,q）） based on genetic algorithm was better than MGM（1,n） model, and the MGM（1,n） model was better than MGM（1,1） model.     

金黄色葡萄球菌SSL7蛋白对补体系统的作用分析

为探究金黄色葡萄球菌超抗原样蛋白(Staphylococcal superantigen-like proteins,SSLs)与补体系统相关的生物学特征,利用原核表达系统表达SSL7重组蛋白,并通过补体杀伤试验、Western Blot以及Pull-down对其有关补体系统的生物学特征进行研究。结果表明:1)重组蛋白SSL7可在大肠杆菌感受态细胞BL21(DE3)系统中大量可溶性表达,大小约25ku;2)SSL7可阻断新鲜兔血清对大肠杆菌DH5α的补体杀伤作用;3)SSL7抑制补体激活后攻膜复合体(Membrane attack complex,MAC)的形成,新鲜兔血清无法在金黄色葡萄球菌NCTC 8325菌体表面形成MAC沉积;4)SSL7能捕获健康人血清(Human normal serum,HNS)中的补体C5,阻断补体系统的激活。综上,金黄色葡萄球菌超抗原样蛋白SSL7可以通过特异性结合补体C5阻断补体级联反应,逃避宿主免疫应答以达到致病的目的,为进一步探究SSL7的感染机制、揭示SSLs蛋白功能奠定基础。     

云芝多糖对奥尼罗非鱼生长、血清溶菌酶活性和补体活性的影响

在基础饲料中分别添加0.5 g/kg、1.0 g/kg、1.5 g/kg、2.0 g/kg、2.5 g/kg的云芝多糖,饲喂(9.0±1.0)g奥尼罗非鱼(rom is niloticus×O.aureus)56 d后,测定罗非鱼增重率、特定生长率、饵料系数、血清溶菌酶活性和血清补体替代途径溶血(ACH50)活力。结果表明:添加量为0.5 g/kg时,试验组罗非鱼增重率和特定生长率显著增高,饵料系数显著降低;添加量为1.0 g/kg时,血清溶菌酶和ACH50活性达到最高,但与对照组差异不显著,而添加量为2.5 g/kg组的血清溶菌酶活性显著低于对照组。     

FMMU白化豚鼠与花色豚鼠血清Ig和补体含量及总补体活性比较

测定比较了 2月龄的封闭群 FMMU白化豚鼠与花色豚鼠的血清免疫球蛋白含量、补体含量及活性。结果表明 :白化豚鼠血清 Ig G含量极显著低于花色豚鼠 ,而 Ig A、Ig M含量显著低于花色豚鼠。白化豚鼠 C3含量极显著低于花色豚鼠 ,C4含量显著低于花色豚鼠。血清总补体活性 ( CH50 )显著低于花色豚鼠。     

Neutralizing and Complement-fixing Monoclonal Antibodies as an Aid to the Diagnosis of Equine Herpesvirus-1 Infection

One complement-fixing (C-MAb) and three complement-dependent neutralizing monoclonal antibodies (N-MAbs) were raised against Hisar-90-7 equine herpesvirus-1 (EHV-1) strain. The target antigen of the C-MAb (2A5) and two of the N-MAbs (1H6, 9C4) was identified as a 140 kDa polypeptide in Western blotting. The target antigen of N-MAb (9C6) could not be identified. Purified polypeptides of five EHV-1 strains isolated from different regions and at different times gave intense bands at 140 kDa when reacted with N-MAb (1H6) in Western blots. In sandwich ELISA, all four MAbs captured the viral antigen from clinical materials, giving a reliable and rapid diagnosis of EHV-1 infection in equines.