江西7个地方鸡品种遗传多样性的微卫星分析

选用30个多态性较好的微卫星标记．检测了丝毛鸟骨鸡、康乐鸡、宁都三黄鸡、余干乌骨鸡、瓦灰鸡、崇仁麻鸡、东乡绿壳蛋鸡等江西省7个地方鸡品种的遗传多样性。利用等位基因频率计算出各群体的平均遗传杂合度（h）、多态信息含量（PIC）、群体间的Nei氏标准遗传距离（Ds）和DA遗传距离。结果表明．30个微卫星位点中有24个微卫星位点在7个鸡群体中的多态信息含量均为高度多态．可作为有效的遗传标记用于鸡品种的遗传多样性和系统发生关系的分析；在7个品种中．杂合度最低的是余干乌骨鸡．为0．6155．杂合度最高的是崇仁麻鸡．为0．6525，各地方鸡品种的杂合度都偏高，反映了群体的遗传多样性非常丰富；7个鸡品种间的DA、Ds遗传距离远近关系一致．但DA距离均比Ds距离大；用UPGMA法对DA和Ds的聚类结果相同．7个鸡品种被聚为3类．余干乌骨鸡、瓦灰鸡、康乐鸡、宁都三黄鸡聚为一类．崇仁麻鸡、东乡绿壳蛋鸡聚为一类．丝毛乌骨鸡独自为一类。     

猪肺炎支原体膜蛋白P46基因在大肠杆菌中的表达

把猪肺炎支原体(Mycoplasma hyopneumoniae)国际标准株232的P46基因克隆进pGEM-T-EASY载体上,通过PCR方法把该基因中三个编码Trp的密码子TGA突变为TGG,然后将该基因亚克隆到载体pMAL-P2X上,得到重组表达载体pMAL-P2X-P46.用该重组载体转化大肠杆菌TB1,得到表达重组菌TB1-pMAL-P2XA-P46,用终浓度为0.3 mM的IPTG在37℃下诱导表达,获得可溶性表达的融合蛋白MBP-P46,在免疫印迹试验中,兔抗MBP高免血清和兔抗猪肺炎支原体高免血清都能与目的蛋白发生阳性反应,证明猪肺炎支原体P46基因在大肠杆菌里获得了可溶性表达.该融合蛋白对于建立特异性和敏感性好的EIISSA方法具有重要意义.     

猪群HEV血清抗体调查及一株新的猪HEV ORF2部分基因序列分析

戊型肝炎病毒(Hepatitis E virus,HEV)主要引起人的戊型肝炎,新近研究发现猪在病毒传播中可能发挥重要作用.本研究对我国部分省区HEV感染血清学调查,在被检的1 138份血清中,有666份(57.5%)为HEV抗体阳性,猪群抗体阳性率随着月龄增长而升高.通过RT-PCR方法从一份猪粪中扩增并克隆了HEVORF2 N端主要抗原决定区339bp基因片段,序列分析显示,该段基因与我国人群HEV基因4型毒株核苷酸序列同源性为85.9%,但氨基酸序列完全一致.这一结果提示我国猪群存在广泛的HEV感染,并在一定程度上与人群HEV毒株密切相关.     

杆状病毒表达SARS冠状病毒纤突蛋白及其抗原性分析

本研究构建了SARS冠状病毒纤突蛋白(S)重组杆状病毒(rBac-SS).SDS-PAGE及Western-Blot分析表明约190Ku左右的重组SARS纤突蛋白(rSS)在rBac-SS感染圆昆虫细胞获得表达,并具有特异免疫反应原性.以rBac-SS感染的昆虫细胞裂解物稀释后直接包被ELISA板,与Vero细胞培养的全病毒裂解物比较,检测SARS-CoV康复病人血清特异抗体,表现出同样的敏感性和特异性;rBac-SS感染的昆虫细胞用于间接免疫荧光,快速检测血清特异抗体反应,具有良好的敏感性和特异性.结果显示,杆状病毒表达的rSS有望替代SARS-CoV全病毒,作为安全、敏感和特异的重组诊断抗原,并为探索重组亚单位疫苗的可行性奠定基础.     

简单、实用、高效的细胞培养技术

通过近几年的实践,笔者对我国兽医生物制品生产中传统的细胞培养工艺进行了一些改进,包括用洗洁精代替其他洗液,用自行研制的营养液代替常规营养液用于细胞培养,改变培养时的转速,改变细胞消化液,改进细胞的保存方法.这些方法均成功应用于生产,取得了良好的效果.     

中草药免疫增强剂对鸡小肠T淋巴细胞免疫组化研究

为了探讨中草药免疫增强剂的作用机理,采用组织学常规切片技术和免疫组织化学染色的方法对鸡小肠黏膜淋巴组织中T淋巴细胞的数量分布进行了动态观察.将1日龄60只健康公鸡随机分成2组,一组为对照组,另一组为试验组,试验组按1%浓度的中草药免疫增强剂饮水,连续21 d.在第14天、第21天分别取各组鸡小肠,制成组织切片,进行组织学观察.结果显示,试验组鸡小肠的T淋巴细胞数显著高于对照组(P<0.05),表明中草药免疫增强剂对鸡小肠免疫有显著的增强作用.     

0～50 cm与0～100 cm土层土壤湿度的转换关系研究

对郑州、南阳2个测点多年来0～100cm土壤湿度、降水(灌溉)及作物发育期资料进行了分析。结果表明,按土壤湿度随时间的变化规律,可将其划分为秋季缓慢下降期、冬季相对稳定期、春季迅速下降期和夏季增墒期4个时段。对0～50cm与0～100cm土层土壤湿度进行回归分析,当前期降水量在10mm以下时,用直线或二次曲线拟合效果良好;当前期降水(灌溉)量大于10mm时,0～50cm与0～100cm土壤湿度之比与前期降水(灌溉)量呈双曲线或二次曲线关系,拟合曲线考虑降水(灌溉)时回归效果有所提高。同一土壤类型、不同时段,或同一时段、不同的土壤类型,拟合的方程不同,反映出不同层次土壤湿度关系的复杂性。     

Effects of gastrin on expression of cyclooxygenase and growth factors in rat gastric mucosa

AIM: To clarify the effects of gastrin on the expression of cyclooxygenase (COX) and several growth factors in rat gastric mucosa. METHODS: Male Sprague Dawley rats were fasted for 24 hours and subcutaneously injected with saline or gastrin 17 at doses of 1 μg/kg, 10 μg/kg and 100 μg/kg, respectively. The expression of COX-1, COX-2, heparin-binding epidermal growth factor-like growth factor (HB-EGF) and hepatocyte growth factor (HGF) in the gastric mucosa were examined using Western blotting and immunohistochemical staining. Effects of a potent gastrin receptor antagonist YM022 on the expression of COX-1, COX-2, HB-EGF and HGF in gastric mucosa were also evaluated. RESULTS: Gastrin dose-dependently increased the expression of COX-2 and HB-EGF in rat gastric mucosa while the expression of COX-1 and HGF did not change significantly after treatment with gastrin. However, pretreatment with YM022 dose-dependently abolished the up-regulation of COX-2 and HB-EGF expression induced by gastrin. CONCLUSIONS: This study demonstrates that gastrin up-regulates COX-2 and HB-EGF expression in rat gastric mucosa, indicating that COX-2 and HB-EGF are involved in pathogenesis of the gastrin-related gastric mucosal hyperplasia and carcinoma of stomach.     

Effects of vasoconstrictor and endothelium-dependent relaxationagent on thoracic aortic rings of diabetes mice

AIM: To discuss the effects of vasoconstrictor and endothelium-dependent relaxation agent on the thoracic aortic rings in different stages of diabetes mellitus. METHODS: Streptozotocin (STZ, 40 mg/kg) was injected intraperitoneally to establish diabetic model in C57BL/6J mice. At the 17th, 22nd, 28th week, diabetic and age-matched control mice were sacrificed respectively and the effects of vasoconstrictors: phenylephrine (PE), 60 mmol/L KCl and endothelium-dependent relaxation agent respectively (ACh) were measured in two groups using thoracic aortic rings. RESULTS: The level of fasting plasma glucose concentration 2 weeks after STZ treatment increased higher (≥11.1 mol/L) in STZ-induced diabetic mice than that in age-matched controls, and maintained at this level in entire experiment course. On the contrary, the weight was decreased significantly. The responses of thoracic aortic rings in STZ-induced diabetic mice to PE were increased, unaltered and increased at the 17th, 22nd, 28th weeks, respectively compared to that in age-matched controls. The responses to 60 mmol/L KCl were also increased in all stages. While the responses to ACh were increased, unaltered and decreased at the 17th, 22nd, 28th weeks, respectively compared to that in age-matched controls. CONCLUSION: The responses of thoracic aortic rings to vasoconstrictor enhance in STZ-induced diabetic mice. However, the endothelial functions potentiate initially due to compensation, and then lower exhibiting endothelial damage.     

Protective effect of ischemic preconditioning on rat retina with sustaining ischemia

AIM: To observe the effect of ischemic preconditioning (IPC) on the subsequent permanent ischemic rat retina. METHODS: A model of two-vessel occlusion (2VO) was used to give an ischemic insult to rat retina. Bilateral carotid arteries were occluded directly in the simple ischemic groups. A procedure of double 2 min ischemia- 3 min reperfusion was applied to IPC groups before their carotid arteries were occluded. Experimental control groups received the same operation except that their exposed vessels were not occluded. Other 6 normal rats served as immunohistochemical controls. Eyeballs were taken out after being subjected to 1, 3 and 7 days of ischemia. The morphometry of retina was measured by stereologic methods. The apoptosis during the retinal ischemia was detected by TUNEL. Immunohistochemistry staining was used for the localization of bcl-2 in retina. RESULTS: In IPC groups, the thickness of retina was thinner than that in simple ischemic groups. The apoptosis was less when compared with the simple ischemic groups at the same ischemic time. The apoptotic cells show only in INL and the apoptosis of RGCs was not appeared until 7 days postischemia. There was no significant difference of the numerical density of RGCs at the different time after the operation. The bcl-2 immunoreactivity was weaker than that of the simple ischemic groups too. CONCLUSION: IPC reduced the injury caused by ischemia in retina, showing a protective effect.