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。     

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。     

固定效应对槟榔江水牛体重影响的研究

[目的]槟榔江水牛为我国目前为止发现的唯一本土河流型水牛,具有产奶性能高、肉用性能好的特点,为了进一步阐明槟榔江水牛的生长发育规律,本文收集云南腾冲县巴福乐槟榔江水牛2004~2011年生长发育数据资料,研究性别、月份和年度等固定效应对体重的影响。[方法]应用SAS分析性别、月份、年度等非遗传因素对槟榔江水牛体重的影响,并且用MTDFREML软件分析了固定效应值。[结果]本研究结果表明,槟榔江水牛性别对初生重具有显著影响,公犊平均初生重(35.04kg)显著高于母犊(32.63kg);月份对母牛初生重没有显著影响,公牛的效应值比母牛的较大。[结论]月份对槟榔江水牛的体重有显著的影响,可能与饲料的季节性供给有关。