Promoter identification and effect on activation of NF-κB of porcine ISG58

Interferon (IFN)-stimulated gene (ISG) 56 family (composed of ISG54, ISG56, ISG58, and ISG60) plays important roles in defense against viral infection in mammalian cells. Numerous studies have been conducted on ISG54, ISG56, and ISG60; however, little is known on ISG58. In the present study, the upstream sequence of porcine ISG58 gene was first characterized as functional promoter by luciferase reporter assay, and then two directly adjacent IFN-stimulated response elements (ISREs), one at ?206 to ?194 (ISRE-I) and a second one, directly upstream of this element at ?219 to ?207?bp (ISRE-II), were identified using the bioinformatics method. The subsequent site-directed deletion and transient transfection experiments showed the candidate ISREs are functional. ISRE-I works better than ISRE-II and synergistic cooperation exists between two ISREs. Additionally, the effect of porcine ISG58 on activation of NF-κB was analyzed using the dual-luciferase reporter assay. The results will contribute to revealing the role of ISG58 in immune response.     

高效液相色谱法测定金荞麦超微粉原花青素B2含量

试验为测定金荞麦超微粉中有效成分原花青素B₂的含量,将四川金荞麦饮片粉碎至0.75 nm以下的超微粉,50%乙醇提取,并利用高效液相色谱测定原花青素B₂的含量。结果显示,在0.0352~1.4080 μg/mL范围内原花青素B₂与峰面积线性关系良好,回归方程为Y=59797X+592.3,r=0.996,金荞麦超微粉中原花青素B₂含量在1.305~1.541 mg/g之间。本试验结果表明该方法能快速、稳定、精确的检测原花青素B₂含量,为金荞麦制剂质量控制提供了科学依据。     

Mucopolysaccharidosis type VI in a Miniature Poodle-type dog caused by a deletion in the arylsulphatase B gene

Abstract

AIM: To investigate and characterise an inborn error of metabolism in a dog with skeletal and ocular abnormalities.

METHODS: A 2.5-year-old small male Miniature Poodle-like dog was presented with gross joint laxity and bilateral corneal opacities. Clinical examination was augmented by routine haematology, serum chemistry, radiographs, pathology, enzymology and molecular genetic studies. Euthanasia was requested when the dog was 3 years of age because of progressively decreasing quality of life.

RESULTS: Radiology revealed generalised epiphyseal dysplasia, malformed vertebral bodies, luxation/subluxation of appendicular and lumbosacral joints with hypoplasia of the odontoid process and hyoid apparatus. These clinical and radiographic findings, together with a positive urinary Berry spot test for mucopolysaccharides, and metachromatic granules in leucocytes, were indicative of a mucopolysaccharidosis (MPS), a lysosomal storage disease. Histological lesions included vacuolation of stromal cells of the cornea, fibroblasts, chondrocytes, macrophages and renal cells. The brain was essentially normal except for moderate secondary Wallerian-type degeneration in motor and sensory tracts of the hind brain. Dermatan sulphate-uria was present and enzymology revealed negligible activity of N-acetylgalactosamine-4-sulphatase, also known as arylsulphatase B, in cultured fibroblasts and liver tissue. A novel homozygous 22 base pair (bp) deletion in exon 1 of this enzyme's gene was identified (c.103_124del), which caused aframe-shift and subsequent premature stop codon. The “Wisdom pure breed-mixed breed” test reported the dog as a cross between a Miniature and Toy Poodle.

CONCLUSIONS: The clinicopathological features are similar to those of MPS type VI as previously described in dogs, cats and other species, and this clinical diagnosis was confirmed by enzymology and molecular genetic studies. This is an autosomal recessively inherited lysosomal storage disease.

CLINICAL RELEVANCE: The prevalence of MPS VI in Miniature or Toy Poodles in New Zealand and elsewhere is currently unknown. Due to the congenital nature of the disorder, malformed pups may be subject to euthanasia without investigation and the potential genetic problem in the breed may not be fully recognised. The establishment of a molecular genetic test now permits screening for this mutation as a basis to an informed breeding policy.     

用单抗研究T细胞与牛流行性白血病发生的关系

本试验对10头患流行性白血病病牛的不同器官的肿瘤组织，用8种单克隆抗体（McAb),TH14B、BAQ44A、PIg45A、BIg715A、PIg501E、cAct105、MM1A、AHCC125染色进行了观察。结果：病牛经临床病理学和免疫琼扩试验而诊断为EBL，通过免疫组织化学检测，在10个病例中发现有9个病例的肿瘤组织对B细胞属性的McAb有很强的染色反应，证明其来源于B细胞。仅有1个病例对B细胞属性的McAb着色很淡，而对T细胞属性的McAb着色很深，呈强阳性反应，说明其来源可能与T细胞有关。     

Prevalence of megabacteria in budgerigar colonies

Objective To measure the prevalence of megabacteria in budgerigar-breeding colonies and to evaluate possible methods to reduce the prevalence.
Design A monitoring study over several years.
Sample population Two budgerigar ( Melopsittacus undulatus ) colonies with over 300 birds each.
Procedure The prevalence of megabacteria in the faeces in two budgerigar breeding colonies, colony 1 and 2, was determined by faecal examination of each bird. Following an initial survey (1990), most of the birds that were scored 2+ or more were culled and a management practice was implemented to discriminate against positive birds. Consecutive yearly surveys (1991, 1992) were conducted on the young birds bred in these colonies. The prevalence of megabacteria in colony 2 was also evaluated in 1994 and 1996 after all the birds were treated with amphotericin B administered in drinking water.
Results The prevalence of megabacteria in the two colonies was significantly (P < 0.001) different. Overall the prevalence of megabacteria adjusted for colony differences was significantly higher (P < 0.025) in males compared to females. Age was not an influencing factor. After the initial survey, the prevalence in the offspring did not significantly (P > 0.05) decrease in the following two annual breeding seasons but by inference it did significantly decrease after amphotericin B treatment.
Conclusion The practice of culling most birds with more megabacteria in faeces and discriminating against positive birds when selecting birds for breeding or culling birds on show quality does not decrease megabacteria prevalence in the offspring. However, a reduction in prevalence does occur with administration of amphotericin B. Birds may have amphotericin B-resistant organisms and these birds need to be identified and culled.     

复方中药提取物在新城疫B_1株免疫中的免疫增强作用

用自制的中药提取物与新城疫Ｂ１株混合后免疫５５日龄鸡,以血凝抑制试验测定新城疫抗体效价。结果表明,５５日龄鸡的中药提取物最佳用量为１．０ｍＬ,试验组鸡ＮＤ抗体效价明显高于对照组。免疫后６０ｄ用Ｆ４８Ｅ８新城疫强毒进行攻击,试验组的保护率达到１００％,而对照组相对较低。     

内质网应激偶联炎性反应研究进展

内质网是真核细胞蛋白合成和折叠的主要细胞器,当内质网内蛋白合成或折叠负担增加,引起未折叠或错误折叠蛋白增多时,可激活内质网几条特定信号通路,启动未折叠蛋白反应,这对维持细胞稳态有重要意义。越来越多研究表明,多种炎性反应疾病与内质网应激有密切联系。一方面,内质网应激引起的未折叠蛋白反应可以诱发或者抑制炎症,另一方面,炎性反应也能影响蛋白折叠,从而促进或缓解内质网应激。2型糖尿病、肿瘤和动脉粥样硬化等多种重大疾病的病理机制都涉及内质网应激与炎性反应的相互作用,对该问题的深入研究不仅能加深人们对这些疾病发病机制的理解,也有助于相关药物的研发。     

Nisin 对 Lactobacillus acidophilus La-5 和 Bifidobac

在MRS平板上，采用琼脂扩散法测定了Nisin对Lactobacillus acidophilus La-5和Bifidobacterium bifidum Bb-12生长的抑制作用。当Nisin浓度≥50ug/ml时，对Lactobacillus acidophilus La-5表现出强烈的抑制作用，而Bifidobacterium bifidum Bb-12则几乎不生长，即使Nisin的浓度仅为25ug/ml，当L.acidophilus La-5和B.bifidum Bb-12单独或两者共同在37℃发酵10％（w/w)还原脱脂奶时，脱脂奶中添加的50ug/ml Nisin对B.bifidum Bb-12表现出杀菌作用，而对L.acidophilus La-5则为抑菌作用。当延长培养时间后。L.acidophilus La-5的存在可以降低Nisin对B.bifidum Bb-12的致死作用。     

检测鸭呼肠孤病毒抗体间接ELISA方法的建立

RT—PCR扩增鸭呼肠孤病毒（Duckreovirus,DRV）的dB基因,克隆到pET-32a（＋）表达载体,再转化大肠杆菌Transetta（DE3）;含有重组质粒pET—σB的大肠杆菌经IPTG诱导,获得大小为55000的以包涵体形式表达的σB重组蛋白。Westernblotting显示,σB重组蛋白能够与兔抗DRV多抗血清特异性结合。以高亲和NI—NTA树脂在变性条件下纯化、梯度尿素复性的σB重组蛋白为包被抗原,建立了检测DRV抗体的间接ELSIA方法。分别以间接ELSIA方法和血清中和试验对DRV感染鸭血清和SPF鸭血清进行检测,两者的符合率为100％。该间接ELSIA方法对鸭瘟病毒、鸭病毒性肝炎病毒和禽流感病毒阳性血清均无交叉反应。