山东PRRSV流行株ORF5、ORF6、ORF7基因序列的分子特征

采用RT-PCR技术对2001~2007年分离自山东地区10株(ShanDong-3、SD-JN、SD-ZQ、SD1、SD2、SD3、SD4、SD5、SD6和SD7)猪繁殖与呼吸综合征病毒(PRRSV)进行ORF5、ORF6和ORF7基因的扩增、克隆和测序,与已知序列的毒株的相应片段进行同源性分析比较,并对其分子特征进行分析。结果表明:该10株病毒仍属北美洲型,其中2001~2002年分离的SD1株、SD2株和2006年分离的SD6株核苷酸序列之间的ORF5、ORF6、ORF7同源性分别为99.2%,99.8%,100%,均与北美洲原型代表株(VR-2332株)和疫苗毒MLVRe-spPRRS Repro USA遗传距离较近,同属一大分支;2006~2007年分离的PRRSV ShanDong-3、SD-JN、SD-ZQ、SD3,SD4,SD5,SD7分离株ORF5、ORF6、ORF7同源性分别为97.8%~100%,99.4%~99.8%,99.2%~99.7%,均与国内96年Ch-1a和2002年HB-1株及2006年分离鉴定的国内高致病性分离株(JXA1、Shanghai、HEB1)同属一个大的分支。首...     

Present Concepts on the Inflammatory Modulators with Special Reference to Cytokines*

The pro- and anti-inflammatory cytokines create a network of interactions between cells that lead to both stimulatory and inhibitory responses that maintain an effective homeostatic regulation. The anti-inflammatory cytokines are a family of peptides that modulate the pro-inflammatory cytokine response. Cytokines act in concert with non-cytokine mediators, such as prostaglandin E₂, glucocorticosteroids, lipocortins, and catecholamines. This review highlights new developments in our understanding of the pathophysiology of inflammation and gives an example of a more recent approach to the modulation of acute systemic inflammatory disorders: activation of ₂-adrenergic receptors on macrophages. In this respect the potent ₂-adrenergic agonist clenbuterol seems of therapeutic interest.     

激活方法对水牛卵母细胞孤雌发育的影响

系统探讨了几种激活方法对水牛卵母细胞孤雌发育的影响。体外成熟 (IVM) 2 6 h的水牛卵母细胞经 7%乙醇(EH)、钙离子载体 (A2 3187)或离子霉素 (Ion)激活处理 5 min后 ,在 2 m mol/ L 6 -甲二氨基嘌呤 (6 - DMAP)中培养 3～4 h,然后再继续培养 6～ 11d,观察其胚胎发育情况。结果发现 ,5μmol/ L Ion激活处理的水牛卵母细胞分裂率显著高于 EH处理的卵母细胞 (6 3.0 % vs5 4 .2 % ,P<0 .0 5 ) ,其原核形成率也显著高于 EH处理的卵母细胞。激活处理前 ,无Ca2 培养液孵育时间的长短 ,对水牛卵母细胞孤雌发育无显著影响 (P>0 .0 5 )。如用 A2 3187激活处理水牛卵母细胞 ,其激活分裂率则随着浓度的升高而提高。从而表明 ,5 μmol/ L Ion可有效激活水牛卵母细胞     

Effects of the Ratio of n-6:n-3 Polyunsaturated Fatty Acids in the Maternal Diet on Immunoglobulins, Fatty Acid Composition, and Performance of Lactating Sows and Suckling Piglets under Conditions of Low Maternal Energy Intake

This experiment was conducted to investigate the effects of different ratios of n-6:n-3 polyunsaturated fatty acids (PUFA) in sow diets on the performance of lactating sows and their piglets at low digestible energy intakes.Twenty-one pregnant Landrace sows were assigned to one of three diets from day 108 of gestation until weaning (26 to 29 d),containing n-6∶n-3 PUFA ratios of 3∶1,8∶1 and 11∶1.The effects on sow and litter production traits and on sow body condition were examined.There were no differences among treatments in daily feed intakes or in changes in sow weight and back-fat levels during lactation.Litter size,litter weight at birth and weaning and litter average daily weight gain were also unaffected by treatment.As expected,large differences were observed in n-6 and n-3 fatty acids in the colostrum and plasma of sows and piglets (P ＜ 0.01 ).The ratio of n-6∶n-3 PUFA the diet was positively correlated with those in colostrum,sow plasma and piglet plasma ( R2 =0.55,0.80 and 0.80,respectively).Sow plasma insulin and IGF-I levels at weaning were unaffected by the treatments.Plasma leptin ( P ＜ 0.05) concentrations were increased in sows fed the diet with a n-6∶ n-3 ratio of 8∶1.Immunoglobulin concentrations in colostrum were not altered by dietary treatment.Plasma IgG concentrations at d 14 were highest in piglets from sows fed the 8∶1 ratio of n-6∶n-3.Furthermore,this group had the highest IgA concentrations at day 21 of lactation compared with the other two groups ( P ＜ 0.05 ).In summary,our study demonstrated that at low digestible energy levels,altering the ratio of n-6∶n-3 PUFA in the diets for lactating sows affected immune components and the fatty acid composition of lactating sows and their piglets.Further studies are needed to examine whether higher levels of fat supplementation than those used in the present study ( 1.5％ ) can successfully enhance performance.     

三肽囊素对鸡血浆蛋白组分及IL-2与IL-6含量的影响

144只1日龄黄羽肉鸡随机分为4组,第7日龄时第Ⅰ～Ⅳ组分别注射0、0.005、0.010、0.030mg/kg剂量的三肽囊素,在14、35及56日龄时每组各取一个重复,取血浆,醋酸纤维薄膜电泳检测血浆蛋白各组分的比率,放射免疫法检测血浆IL-2与IL-6的含量.结果显示不同剂量的三肽囊素可提高血浆γ球蛋白水平:对血浆...     

肠道病毒71型感染C57BL/6小鼠模型的初步建立

以1日龄C57BL/6乳鼠作为试验对象,分别通过颅内和腹腔注射肠道病毒71型(EV71)福建株感染,观察记录小鼠体重和体征,定期采集小鼠大脑、小肠、肺、肌肉4种组织,分析病毒载量,通过HE染色和免疫组化观察各种组织病理变化。结果:2种感染途径的C57BL/6乳鼠均于不同时间出现竖毛、弓背和消瘦症状。经腹腔注射感染的乳鼠能在肌肉组织中检测到病毒RNA,经颅内注射感染的乳鼠可在肌肉和肺组织中检测到病毒RNA。病理学及免疫组化结果显示:EV71福建株在骨骼肌中大量增殖,并可导致肌肉组织水肿和坏死、肺组织水肿充血以及神经元坏死等多种组织器官炎症反应。综上,初步建立通过颅内和腹腔注射感染EV71的C57BL/6乳鼠模型,为研究EV71病毒感染机制、建立更优的动物模型提供参考。     

鸡白细胞介素-6原核表达及双抗夹心ELISA方法的建立

应用分子生物学技术原核表达ChIL-6,以ChIL-6重组蛋白为免疫原,按免疫程序分别制备兔抗和鼠抗IL-6重组蛋白的多克隆抗体。应用此抗体建立双抗夹心EL-ISA方法后,为了优化此方法本试验对该方法的最佳试验条件、标准曲线、重复性和初步应用进行确定。结果显示,经SDS-PAGE和Western-blot分析,表明ChIL-6在大肠杆菌中正确表达,为了建立检测ChIL6的双抗夹心ELISA方法,本试验应用表达的重组蛋白制备兔抗和鼠抗多克隆抗体。建立的双抗夹心ELISA方法最佳反应条件为包被抗体的质量浓度为50mg／L,4℃过夜;酶标二抗稀释度为1：400,37。C1h;应用该方法检测感染金黄色葡萄球菌后的ChIL-6,其结果与本实验室之前应用人的ELISA试剂盒检测的结果相似。结果表明,本试验建立的检测ChlL-6的双抗夹心ELISA方法可用于临床。     

Tumour necrosis factor stimulated gene 6 intrinsically regulates PD-L1 expressions in breast cancer cells,leading to modulation of tumour microenvironment

Recent studies have shown that tumour cells express tumour necrosis factor-inducible gene 6 (TSG-6) and its protein, which is known to play a key role in regulating excessive immune responses and proliferation and growth of mesenchymal stem cells (MSCs). It has not been confirmed whether the inhibition of TSG-6 for tumour cells can suppress tumour cell growth and regulate the activation of immune cells in the tumour microenvironment (TME). TSG-6-specific small interfering RNA was transfected into canine and human breast cancer cells (CIPp, CIPm and BT-20). TSG-6-down-regulated (siTSG-6) cells showed decreased cell proliferation, migration, and invasion abilities. Decreased mRNA expressions of NF-κB, STAT3 and Sox2, confirming that TSG-6 is an upper factor governing tumour growth and metastasis. Notably, siTSG-6 cells showed significantly decreased expression levels of CD44 and PD-L1. Direct and indirect co-culture of canine peripheral blood mononuclear cells (cPBMCs) and the siTSG-6 cells showed significant activation in M1 type macrophages and cytotoxic T cells. They also showed a tendency to decrease in the expression of CTLA-4 and increase in the expression of PD-1. In conclusion, this study suggests that the down-regulation of TSG-6 in breast cancer cells could not only suppress tumour growth and metastasis, and but also regulate TME. Since modulation of immune checkpoint proteins occurs in both tumour cells and immune cells, inhibiting TSG-6 and its protein within the TME could be novel therapeutic target for anticancer treatment.     

Wnt10b基因的生理功能及其在绒毛周期性生长中的作用研究进展

绒山羊的绒毛生长具有周期性的特点,受多种因素的影响。Wnt10b基因作为Wnt家族的重要成员之一,在动物体脂肪的生成、骨骼的生成和某些疾病（如肿瘤）的产生、毛囊的生成,以及毛发基板的生长与保持活性状态等方面发挥着重要作用。对Wnt10b的来源、结构、生理功能、调控机制及其对毛囊周期性生长的影响进行综述,以期为Wnt10b促进绒毛周期性生长的作用机理研究提供参考。