牛GM-CSF与金黄色葡萄球菌黏附素FnBPB共表达质粒的构建与原核表达

采用PCR方法对牛粒细胞-巨噬细胞集落刺激因子(GM-CSF)和金黄色葡萄球菌FnBPB的D区进行特异性的扩增,并通过重叠延伸PCR扩增GM-CSF-FnBPB串联基因,构建了克隆质粒pMD19-GM-CSF-FnBPB。利用表达载体pET-32a(+)对该融合基因片段进行原核表达,SDS-PAGE分析表明,在1mmol/L IPTG诱导浓度下,在39ku处出现了与目的蛋白一致的外源蛋白带,Western blot分析表明,该蛋白具有反应原性,进而证明该融合基因成功在原核细胞中表达。     

奶牛免疫功能的影响因素

影响奶牛免疫功能的因素复杂,围产期、能量负平衡以及饲料蛋白质、维生素、某些微量元素的缺乏会引起的奶牛出现免疫功能的降低,饲料中添加某些维生素、微量元素、壳聚糖、酵母细胞壁等添加剂可以增强奶牛的免疫功能,此外,某些中药也可增强奶牛的免疫功能。     

山羊成纤维生长因子受体（FGFR1）基因的多态性检测

选择与羊同源性较高的牛FGFR1基因组序列（NM_001110207）设计3对特异性引物,采用DNA池PCR直接测序法快速检测黔北麻羊FGFR1基因的多态性,并以内蒙古白绒山羊和关中奶山羊比较,结果表明：在引PCR产物中只有黔北麻羊检测到多态,分别处于外显子5和内含子5的T133C和C211T的碱基突变,而其他2个品种...     

Prevalence and risk factors associated with intestinal parasites in pigs in Chongqing, China

From 2007 to 2009, the prevalence of intestinal parasites was investigated in intensive and extensive pig farms in Chongqing, China. A total of 2971 samples from both sexes and five age categories (breeding boars, breeding sows, fatteners, growers and weaners) were evaluated by standard methods for the presence of helminth ova and protozoan oocysts, cysts and/or trophozoites. Of the 2971 pigs sampled, 362(12.18%) were infected with Ascaris suum, 301(10.13%) with Trichuris suis, 301(10.13%) with Oesophagostomum spp., 491(16.53%) with Eimeria spp., 149(5.02%) with Isopora suis, 677(22.79%) with Balantidium coli and 196(6.60%) with Cryptosporidium spp. Growers had the highest infection rate while breeding boars had the lowest among the five age categories. B. coli was the most common protozoan in all pig age groups. Pigs infected with multiple parasites were common. Risk factors such as management methods, seasons, ages, etc. can influence the infection rate to a certain degree. This investigation provides relevant data about risk factors for pig farmers, thus allowing them to make more appropriate antiparasitic treatments according to farm conditions and local climate in Chongqing.     

Low-density lipoprotein-related receptor protein 1 (LRP-1) is not required for insulin-like growth factor binding protein 3 (IGFBP-3) to suppress L6 myogenic cell proliferation

Insulin-like growth factor binding protein-3 (IGFBP-3) suppresses proliferation of numerous cell types, including myogenic cells, via both insulin-like growth factor (IGF)-dependent and IGF-independent mechanisms; however, the mechanism of IGF-independent suppression of proliferation is not clearly defined. In nonmuscle cells, binding of IGFBP-3 to the low-density lipoprotein receptor-related protein-1 (LRP-1)/activated α(2)M receptor is reportedly required for IGFBP-3 to inhibit proliferation. These findings suggest that binding to this receptor also may be required for IGFBP-3 to suppress proliferation of cultured myogenic cells. To investigate the role of the LRP-1 receptor in suppression of myogenic cell proliferation by IGFBP-3, we have examined the effect of receptor-associated protein, an LRP-1 receptor antagonist, on recombinant porcine (rp)IGFBP-3 inhibition of L6 myogenic cell proliferation. Treatment with receptor-associated protein results in a 37% decrease (P < 0.05) in the ability of rpIGFBP-3 to inhibit L6-cell proliferation. In L6 cells subjected to LRP-1 small interfering RNA treatment for 48 h (LRP-1 silenced), LRP-1 mRNA levels were reduced by greater than 80% compared with control cultures treated with nonsense small interfering RNA (mock silenced). In addition, the 85-kDa transmembrane subunit of LRP-1 was undetectable in Western immunoblots of total protein lysates from LRP-1-silenced cells. Even though LRP-1 mRNA and protein levels were dramatically reduced in LRP-1-silenced L6 cells compared with mock-silenced controls, rpIGFPB-3 suppressed proliferation rate to the same extent in both LRP-1-silenced and mock-silenced cultures. Our results strongly suggest that, in contrast to data obtained for nonmuscle cell lines, the LRP-1 receptor is not required for IGFBP-3 to suppress proliferation of L6 myogenic cells.     

Polymorphism in promoter region of growth hormone receptor is associated with potential production capacity of insulin‐like growth factor‐1 in pre‐pubertal Holstein heifers

Insulin‐like growth factor‐1 (IGF‐1) is one of the important factors for growth, milk production and reproductive functions and mainly released from the liver in response to growth hormone (GH) via GH receptor (GHR) in cattle. Recently, some single nucleotide polymorphisms (SNPs) were identified in the bovine GHR gene. Some GHR‐SNPs were shown to be related to plasma IGF‐1 concentration in cattle. Hence, the capacity to IGF‐1 production in the liver might be affected by GHR‐SNP and associated with performance in the future. This study examined whether GHR‐SNP is associated with IGF‐1 production in the liver of pre‐pubertal heifers. In 71 Holstein calves, blood samples for genomic DNA extraction were obtained immediately after birth. To genotype the GHR‐SNPs in the promoter region, polymerase chain reaction (PCR) products were digested with restriction enzyme NsiI (cutting sites: AA, AG and GG). All heifers at 4 months of age were intramuscularly injected with 0.4 mg oestradiol benzoate. Blood samples were obtained from the jugular vein just before (0 h) and 24 h after injection. The number of AA, AG and GG at the NsiI site was 0, 17 and 54 respectively. In AG and GG, plasma GH concentrations were higher pre‐injection than 24 h post‐injection (p < 0.01). Moreover, plasma GH concentrations in AG post‐injection were higher than in GG (p < 0.05). In contrast, the GG genotype exhibited higher plasma IGF‐1 concentrations in pre‐injection than post‐injection (p < 0.01), although oestradiol did not change IGF‐1 concentration in the AG genotype. We conclude that the GG polymorphism in the promoter region of GHR is associated with a higher potential capacity of IGF‐1 production in the liver of cattle.     

控制sch单养雄蚕品种雌雄蚕孵出比例的催青处理因素研究

研究结果表明 :在RH 6 0 %状态下 ,控制性比的温度和下限时间负相关达到显著水平 ;控制性比的高温处理适于干燥状态下进行 ;处理后雌蚕孵化整齐度明显低于雄蚕孵化整齐度 ,雌雄蚕孵化整齐度均随处理温度的升高而下降 ,尤以雌蚕更为明显。由此提出了在RH 6 0 %状态下sch单养雄蚕品种控制性比的起点温度及温度、时间范围。认为高温催青用下限温度配合 1日收蚁法效果好 ,高温处理以后的胚胎适于在RH 80 %～ 85 %的湿度中保护     

GDNF对海仁酸损伤体外培养新生大鼠背根神经节神经元的影响

N1无血清培养基原代分离培养新生大鼠背根神经节（dorsal root ganglion DRG)神经元，用兴奋性毒素海仁酸（kainic acidKA)损伤体外培养细胞后，加入胶质源性神经营养因子（glial-derived neurotrophic factor,GDNF)作用。最后通过MTT法检测，细胞总蛋白测定，胎盘兰染色计数，形态学观察等方法研究分析GDNF对兴奋性毒素海仁酸损伤后大鼠DRG神经元的活性，存活及突起生长的影响，结果表明：GDNF对体外正常生长的DRG神经元的存活，活性及突起生长没有明显的促进作用。而在海仁酸损伤后，GDNF对其存活和活性有明显的促进作用。但对突起生长没有明显的影响。