鸡粒细胞巨噬细胞集落刺激因子-猪干扰素α1融合基因在大肠埃希菌中的可溶性表达及其生物学活性研究

为可溶性表达重组鸡粒细胞巨噬细胞集落刺激因子(ChGM-CSF)与猪干扰素α1(PoIFNα1)融合蛋白,并研究其生物学活性,本研究分别提取鸡、猪肝脏细胞总RNA,采用RT-PCR方法扩增ChGM-CSF和PoIFNα1基因,经linker连接上述两种基因后将其克隆于pET-32a原核表达载体,转化E.coliBL21(DE3)菌株进行诱导表达,SDS-PAGE和western blot检测融合蛋白表达产物。在ST细胞/VSV病毒测定系统以细胞病变抑制法滴定该融合蛋白的抗病毒活性;同时用MTT法检测其对鸡淋巴细胞增殖活性的促进作用。结果显示,PCR扩增并融合后的ChGM-CSF和PoIFNα1融合基因约为1000bp,构建重组表达载体后,诱导表达的rChGM-CSF-PoIFNα1融合蛋白分子量约55ku,主要存在于破碎菌体的上清中,表达量较高。Westernblot检测结果显示,该融合蛋白分别能够与ChGM-CSF多抗和PoIFNα单克隆抗体特异性结合,其抗病毒比活性约为1.1×10^6IU/mg,并且具有促进鸡淋巴细胞增殖的活性。本研究为rChGM-CSF-PoIFNα1重组融合蛋白的研制及其相关活性的测定提供实验依据。     

Acute phase proteins and biomarkers of oxidative status in feline spontaneous malignant mammary tumours

Acute phase proteins (APP) and biomarkers of oxidative status change in human and canine mammary tumours, however, they have not been studied in feline mammary tumours. The aims of this study were to investigate the APP and antioxidant responses in feline malignant mammary tumours, to evaluate their relation with tumour features, and to assess their prognostic value. Serum amyloid A (SAA), haptoglobin (Hp), albumin, butyrylcholinesterase (BChE), insulin‐like growth factor1 (IGF1), paraoxonase1 (PON1), total serum thiols (Thiol), glutathione peroxidase (GPox) and total antioxidant capacity determined by different assays, including trolox equivalent antioxidant capacity assessed by two different methodologies (TEAC1/2), ferric reducing ability of plasma (FRAP), and cupric reducing antioxidant capacity (CUPRAC), were determined in serum of 50 queens with spontaneous mammary carcinomas and of 12 healthy female cats. At diagnosis, diseased queens presented significantly higher SAA and Hp, and lower albumin, BChE, GPox, TEAC1, TEAC2 and CUPRAC than controls. Different tumour features influenced concentrations of APP and antioxidants. Increases in serum Hp, and decreases in albumin, Thiol and FRAP were significantly associated with neoplastic vascular emboli, metastasis in regional lymph nodes and/or in distant organs. Distant metastasis development during the course of the disease was associated with increases in SAA and TEAC1. At diagnosis, decreased albumin was associated with a longer survival, and BChE <1.15 μmoL/mL.minute was associated with a shorter survival time on multivariate analysis. Feline malignant mammary tumours are associated with an APP response and oxidative stress, and different tumour features influence the inflammatory response and the oxidative damage. Furthermore, some of these analytes proved to have prognostic value.     

青藏高原放牧高寒草甸主要温室气体通量及其主控因素研究

在青藏高原高寒草地不同放牧强度区设4个试验样地,分别为原生草甸(NM)、轻度退化草甸(LM)、中度退化草甸(MM)和重度退化草甸(HM),监测草地温室气体通量。结果表明:不同放牧强度对草地温室气体通量影响显著,放牧活动对高寒草甸的影响首先表现在植被上,而土壤环境的变化比较迟滞。通过逐步回归分析和因子拆分得知,草甸甲烷通量影响较大的环境因素为土壤紧实度和有机质,分别能解释44.6%和28.4%的总变异,CO2通量影响较大的环境因素为紧实度和生物量,分别能解释36.1%和32.8%的总变异,氧化亚氮通量影响较大的环境要素为紧实度和有机质,分别能解释50.1%和22.9%的总变异,家畜的践踏作用使退化草地紧实度增加,进而引起温室气体通量的改变,高寒草甸退化演替发展到重度退化阶段时释放大量温室气体。     

Summary of Ammonia and Particulate Matter Emission Factors for Poultry Operations

Limited data on pollutant emissions from poultry operations are available to assess the effect of these operations on the environment and to put their contribution in perspective with other sources of pollutants. To alleviate this problem, numerous studies at various poultry facilities have been undertaken to improve the knowledge base in quantifying emissions of NH₃ and size-fractionated particulate matter (PM). For these emission data to be of practical use for government agencies and policy makers, the emission rates must be reported as an emission factor with a production unit that enables the emissions from one poultry operation to be correlated to another poultry operation. This paper presents a compilation of NH₃ and PM emission data from several studies in the form of emission factor on a per-500 kg of live weight or animal unit basis. In addition, best management practices that lower pollutant emissions from poultry operations have been reported along with their effectiveness at reducing NH₃ and PM. Unfortunately, the compiled data were insufficient to characterize the variability in emissions caused by differences in house design, suggesting that more studies are needed to complete a comprehensive emission inventory. Once complete, this inventory will enable poultry producers to estimate emissions from their facilities and, if necessary, select management practice(s) to lessen their emissions of NH₃, PM, or both.     

穿心莲超声提取工艺研究

优选穿心莲药材的提取工艺。通过单因素和正交试验,测定穿心莲内酯、脱水穿心莲内酯、新穿心莲内酯三种二萜内酯类的总含量,确定最佳超声提取工艺。确定穿心莲药材的最佳超声提取工艺为乙醇浓度60%,超声时间1h,料液比为1∶15,此条件下穿心莲二萜内酯类的总含量为2.27%。该试验确定的超声提取工艺稳定可行,方法可靠,可为穿心莲有效成分的提取提供理论依据。     

日本结缕草ZjNAC1的克隆、转录激活活性、亚细胞定位及表达分析

采用RACE技术从日本结缕草中克隆得到ZjNAC1转录因子(GenBank No. MH428376),其开放阅读框为945bp,编码314个氨基酸。ZjNAC1编码的蛋白在N端有1个典型的NAM保守结构域,属于NAC转录因子家族。通过染色体步移的方法,获得ZjNAC1基因ATG上游1635bp序列,分析发现其包含响应脱落酸(ABA)、茉莉酸甲酯(MeJA)及逆境胁迫的作用元件。构建pGBKT7-ZjNAC1载体,转化Y2HGold酵母感受态细胞,发现ZjNAC1具有转录激活活性。农杆菌介导的35S-ZjNAC1-YFP载体注射本生烟草叶片瞬时表达结果显示,ZjNAC1定位于细胞核。实时荧光定量结果表明,ZjNAC1在日本结缕草根中表达量最高;此外,ZjNAC1的表达受10μmol/L MeJA和20%PEG4000处理的诱导,但受200μmol/L乙烯(ET)、10μmol/L ABA和300mmol/L NaCl处理的抑制。     

Effect of genistein on IGF‐1 and IGFBP‐1 in young and aged female rat ovary

The objective of this study was to assess the effects of genistein (GEN) on expression of insulin‐like growth factor 1 (IGF‐1) and insulin‐like growth factor binding protein 1 (IGFBP‐1) in young and aged rat ovary. Forty young female Sprague Dawley (SD) rats (200 ± 20 g) and forty aged female SD rats (490 ± 20 g) were selected and according to weight, they were divided into the following five groups with eight animals in each: negative control group (NC), low‐dose group (L), middle‐dose group (M), high‐dose group (H) and positive control group (PC). GEN group received GEN of 15, 30, 60 mg/kg respectively. It lasted 30 days. Concentrations of serum hormones, IGF‐1 and IGFBP‐1 were determined by enzyme‐linked immunosorbent assay (ELISA). Gene and protein expressions of IGF‐1 and IGFBP‐1 were determined by real‐time PCR and Western blot respectively. Compared with NC, GEN significantly increased oestradiol‐17β(E₂) level in aged rat, reduced luteinizing hormone (LH) level in young and aged rat. Serum levels of IGFBP‐1 in young rats were significantly higher in GEN groups (p < 0.05). mRNA and protein expression levels of IGF‐1 and IGFBP‐1 were positively correlated with GEN dose. GEN could significantly reduce the ratio of IGF‐1/IGFBP‐1 of aged rats. Multivariate Cox regression analysis result showed IGF‐1 and IGFBP‐1 levels significantly correlated with GEN dose. We speculate that there is an association between the addition of GEN and expression of IGF‐1 and IGFBP‐1, and the relationship between them is different in young and aged rat.     

我国不同区域鸡源大肠杆菌的致病性与耐药性状分析

为比较我国不同地区发病鸡源大肠杆菌的致病类型、毒力因子差异和其耐药状况,探讨大肠杆菌对家禽生产和公共卫生的影响,对从山东、西藏等7个省(自治区)的发病鸡群分离获得的130株大肠杆菌菌株,用PCR方法进行了肠致病性大肠杆菌、肠毒素型大肠杆菌等5种致病类型检测和ompT、stx2等7种毒力因子检测,并用15种药物测其耐药性。结果发现,仅5株发病鸡源大肠杆菌为致病性大肠杆菌(3.85%,5/130),且均为肠产毒性大肠杆菌(ETEC)。130株大肠杆菌检测出iroN毒力因子检出率最高(69.23%,90/130),stx2最低(1.54%,2/130),其余均在50%~60%。分离菌株对四环素和氨苄西林耐药率最高,分别为91.54%(119/130)、90.77%(118/130),对美罗培南耐药率最低(12.31%,16/130);多重耐药严重,94.62%(123/130)的菌株对三类或三类以上的药物耐药。本研究通过比较不同地区养鸡场大肠杆菌的致病性和耐药情况,以进一步做好大肠杆菌病的防控。