O型口蹄疫病毒VP1 T细胞与B细胞表位基因双拷贝串联表达产物的免疫应答

以一株O型口蹄疫病毒(FMDV)外壳蛋白VP1基因为模板,合成与细胞免疫及体液免疫相关抗原表位肽基因:21-40肽(20AA)和141-160肽(20AA)基因序列,运用基因工程技术构建了含有串联结构21-40(20AA)～141-160(20AA)～21-40(20AA)～141-160(20AA)的2020-2020VP1融合基因表达载体r2020-2020,转化宿主菌BL21(DE3)RIL后诱导表达,表达产物经SDS-PAGE及Western Blot分析显示重组融合蛋白的分子量约为18Ku.动物实验表明,较小剂量的融合蛋白就能诱导豚鼠产生特异性T淋巴细胞增殖反应及抗FMDV中和抗体,证明该融合蛋白可同时激活细胞免疫及体液免疫反应,具有开发成为抗FMDV疫苗的应用价值.     

恩诺沙星在猪组织中残留消除规律研究

在常规饲养条件下,对健康猪按2.5 mg/kg体重的剂量肌肉注射2.5%恩诺沙星注射液,每日2次,连续注射3 d.停药后第2、4、6、8、10、12、14天分别屠宰4头猪.分别采取每头猪的肌肉(注射部位)、脂肪(腹脂)、肝和肾脏等4种组织,用高效液相色谱法进行残留量测定.结果表明:残留在肌肉、脂肪组织中的药物消除较快,第8天总残留量(恩诺沙星 环丙沙星)已下降至检测限(20μg/kg)以下;肝和肾脏组织中的药物消除缓慢,第14天测得猪肾中药物总残留量为40μg/kg.     

新兽药喹烯酮亚慢性经口毒性研究

研究了新兽药喹烯酮对大鼠的亚慢性毒性作用.SD大鼠连续6个月经口染毒,剂量分别为32.8、164、820 mg/(kg bw·d),观察染毒期间大鼠一般情况、体重和进食量,于染毒第3和第6个月进行血常规和生化指标检测,取染毒第6个月大鼠的尿分析尿常规指标;染毒结束时处死全部实验动物,计算脏器系数,进行组织病理检查并分析喹烯酮对脏器的损害情况.研究结果表明,820 mg/(kg bw·d)剂量组雄、雌性动物分别有2、3只死亡,且动物的体重明显低于对照组,脑、肺、肝、脾等主要脏器的脏器系数及雄性动物总蛋白、白蛋白含量均显著高于对照组,病理组织学检查发现肝胆管增生;164 mg/(kg bw·d)剂量组雄性大鼠肝、脾、睾丸和雌性大鼠脑、心、肝脏的脏器系数显著高于对照组.因此,新兽药喹烯酮对大鼠的最大无作用剂量为32.8 mg/(kg bw·d).     

转基因植物疫苗研究进展

众多研究表明,病毒的抗原决定簇及细菌毒素亚单位均能在转基因植物中成功表达,并保持良好的免疫原性.与现有的疫苗生产体系相比,植物生产疫苗具有安全、经济、稳定、高效等优势.本文概述了利用不同受体植物生产疫苗的研究现状、免疫原性评价及免疫原基因的优化表达策略.     

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.     

小麦干湿交替条件下的水分利用效率与生物学性状

在防雨棚盆栽试验条件下,以“A115”、“4185”为材料,用干湿交替方法研究了3种水分组合下小麦的水分利用效率与生物学性状的相关关系。结果表明产量水平的水分利用效率与株高、穗数、单株生物量、光合速率、蒸腾速率和产量成显著正相关;与除此而外的其他被测生物学性状不相关或相关不显著。叶片水平的水分利用效率与气孔导度和蒸腾速率成极显著负相关关系,与穗粒数和分蘖数成显著负相关关系;与其他被测性状不相关或相关不显著。     

High density lipoprotein subclasses and activities of protein kinase C

AIM: To investigate the relationship between high density lipoprotein (HDL) subclasses and the binding activities of hepatic cell and extra-hepatic cell HDL receptor and intracellular protein kinase C (PKC). METHODS: Using purified pre-β1 HDL and apoE-deficent HDL3 to react with human smooth muscle cells (SMC) and HepG2 cells, then the activities of pre-β1HDL and apoE-deficent HDL3 binding to SMC and HepG2 cells, and the effects of this two HDL subclasses on PKC activities in SMC and HepG2 cells were observed . RESULTS: The results showed that the value of Kd of pre-β1HDL binding to SMC HDL receptor was not only significantly lower than that of apoE-deficient HDL3 (P<0.05), but significantly lower than pre-β1 HDL binding to HepG2 HDL receptor (P<0.05). Cell PKC system was all activated by binding of the two HDL subclasses with SMC HDL receptor, and the effect of pre-β1HDL appeared to be stronger than apoE-deficent HDL3. No change in HepG2 cell PKC activity by binding the two HDL subclasses with HepG2 HDL receptor was observed. CONCLUSIONS: The results indicate that the ability of pre-β1HDL to promote efflux of cholesterol from extra-hepatic cells is stronger than that of apoE-deficent HDL3, and it seems that plasma pre-β1 HDL mainly interacts with extra-hepatic cell HDL receptor, subsequently, activates PKC signal transduction system and promotes the intracellular cholesterol efflux from cells.     

Effect of L-carnitine on apoptosis and oxidant damage of cardiomyocytes induced by hypoxia/reoxygenation in vitro

AIM: To examine the effects of L-carnitine on apoptosis and oxidant injury in cultured neonatal rat cardiomyocytes induced by hypoxia/reoxygenation and its possible mechanism. METHODS: The cultured cardiomyocytes were divided into three groups， control， A/R group （anoxia for 120 min, reoxygenation for 240 min） and L-carnitine treatment group, in which cells were exposed to 20 mg/L, 50 mg/L, 100 mg/L, 200 mg/L L-carnitine respectively at 2 h before anoxia. The superoxide dismutase (SOD), succinate dehydrogenase (SDH) activities and malondialdehyde (MDA) content were examined, and the apoptosis was determined by flow of cytometry (FCM). In addition, the ultrastructure was observed by transmission electron microscopy. RESULTS: In A/R group, SOD and SDH activities were lower, the apoptosis rate and MDA content were higher than those in control group (P<0.05). In L-carnitine treatment group, SOD and SDH activities were higher, the apoptosis rate and MDA content were lower than those in A/R group, a L-carnitine concentration-dependent effect was found. Moreover, impairment of myocardial ultrastructure was more severe in A/R group than L-carnitine treatment group. CONCLUSION: L-carnitine can protect cardiomyocytes against hypoxia/reoxygenation-induced injury in a dose-dependent manner.     

Role of PKC and MMPs in the pathogenesis of the rat experimental atherosclerosis and the action of Danshen injection

AIM: To study the effects of protein kinase C on the expression of MMPs that may play an important role in the formation of atherosclerosis and the possible mechanism of Danshen injection to treat atherosclerosis. METHODS: 50 Sprgue-Dawley (SD) rats were divided into three groups randomly: control group (group C), model group (group M) and Denshen treatment group (group D). The serum was collected to measure the level of cholesterol, triglyceride, low density lipoprotein cholesterol（LDL-ch）. The expression of PKC and MMPs were measured by immunohistochemistry. Light microscope and electron microscope were also used. RESULTS: ① The cholesterol, triglyceride, LDL-ch concentrations in group M and group D were significantly higher than those in group C (P<0.01), cholesterol, triglycerideand LDL-ch contents were lower in group D than group M (P<0.01). ② The expression of PKC, MMP-2, MMP-9 in group M were significantly higher than those in group C (P<0.01), and were significantly lower in group D than those in group M (P<0.01). The expression of PKC was correlated with MMP-2 and MMP-9. ③ Light microscope showed that there were plaques in intima and serious calcification, necrosis, obviously irregular thickness in media of group M, but slightly in group D. ④ Electron microscope showed that smooth muscle cells of group M were necrosed , collage grew abundantly and alined disorderly, most of the endothelial cells exfoliated, but slightly in group D. CONCLUSIONS: ① MMPs play an important role in the generation and development of atherosclerosis. ② The activation of PKC may take part in the formation of atherosclerosis and it may be the upstream mechanism of the expression of MMPs. ③ The reduced expression of MMPs and PKC is a part of mechanism for Danshen to prevent and treat atherosclerosis.     

Study on the activation of blood platelets by propylene-acidamide grafted polypropylene membrane in vitro

AIM: To evaluate the blood compatibility of a new bioartificial reactor membranous material (propylene-acidamide grafted polypropylene membrane, PP-g-AAm) in vitro. METHODS: Contacted PP-g-AAm membrane and PP (polypropylene) membrane with platelet-rich plasma in a swing bed, 37 ℃, to simulate the conditions in vivo, and another group of PRP without any membranes was set as control group. ELISA was used to study the expression of β-thromboglobulin, and flow cytometry was used to study CD62P and CD63 expression of the activated blood platelets after contacting the two kinds of membranes with PRP. Scanning electrical microscopy was used to study the configuration and numbers of platelet cells adhered on the membranes. RESULTS: After contacting with PRP 30 min, β-TG expression showed marked difference between the two kinds of material groups and the control group (P<0.01, P<0.05), and the difference between the two kinds of membrane groups was also significant (P<0.05). There were obviously differences on the expression of CD62P and CD63 between the two kinds of membranes after contacting with PRP for 30 min (P<0.05，P<0.01). When enlarged 10 000 times, the disfiguration of the platelet cells adhered on the two kinds of membranes after one hour were found by scanning electrical microscopy, and the numbers of platelets on the PP membrane were more than the PP-g-AAm membrane markedly. CONCLUSION: The PP-g-AAm membrane has better blood compatibility than the PP membrane.