鸡传染性贫血病毒VP1、VP2基因的表达及免疫原性研究

为探索鸡传染性贫血重组蛋白亚单位疫苗的可行性,将鸡传染性贫血病毒基因分别克隆到转移载体p Fast Bac HTA中,再将其分别转化DH10Bac感受态细胞得到相应的重组表达载体,转染昆虫细胞Sf21获得含有VP1、VP2基因的重组杆状病毒v Bac-VP1、v Bac-VP2,应用悬浮培养的Sf21细胞表达重组蛋白。SDS-PAGE及Western blotting结果证明,VP1、VP2基因在昆虫细胞中得到了表达,动物试验进一步证实,重组蛋白免疫SPF鸡可产生ELISA抗体,提示杆状病毒表达的VP1、VP2具有良好的免疫原性。     

用PCR方法检测猪圆环病毒Ⅱ型

建立了一种能特异性地扩增猪圆环病毒 型 (porcine circovirus type 2 ,PCV- 2 )核酸片段的 PCR方法 ,从疑似断奶猪多系统衰竭综合征 (PMWS)的感染病例的淋巴结和脾脏中扩增出了预期长度的 PCV- 2 DNA片段 ,用限制性内切酶 Eco R 酶切鉴定了 PCR产物的特异性。 PCR产物的核苷酸序列分析表明 ,与已报道的 PCV- 2 (Gen Bank Ac-cession num ber AF0 2 72 17)相关区域的核苷酸序列同源性均大于 96 %。首次证实了我国猪群中存在 PCV- 2感染的PMWS。     

胆酸负荷对妊娠后期绵羊母体和胎儿血浆IGF—2及其代谢物水平的影响

以绵羊及其胎儿为试验对象，研究了胆酸负荷对妊娠后期母体和胎儿血浆胰岛素样生长因子－2(in-sulin-like growth factor-2,IGF-2）及其代谢物水平的影响。将7只确知妊娠时间的绵羊分为试验组5只和对照组2只，试验前装置颈静脉瘘，在妊娠90h和104d2闪给试验组静脉灌注胆酸盐1．1mg/kg，给对照组静脉灌注等量生理盐水，妊娠120d时手术安装胎儿后肢胫前动脉血管瘘。每天采胎儿和母体的血样，分离血浆，测定血浆IGF－2和主要代谢物水平。同时测定胎儿出生时生长发育指标。结果表明：（1）试验组母羊IGF－2水平低于对照组，相反其胎儿IGF－2水平却高于对照组（P＜0．05）；（2）试验组母羊的总蛋白和乳酸水平分别比对照组35．64％；（3）试验组胎儿出生重比对照组小29．00％，胎龄短7．50d。因此认为，胆酸重复灌注可引起妊娠绵羊早产和胎儿出生重降低，而这可能与IGF－2和主要代谢物水平变化有关。     

Detection of γ-H2AX,a Biomarker for DNA Double-strand Breaks,in Urinary Bladders of N -Butyl- N -(4-Hydroxybutyl)-Nitrosamine-Treated Rats

To evaluate the potential role of DNA repair in bladder carcinogenesis, we performed an immunohistochemical analysis of expression of various DNA repair enzymes and γ-H2AX, a high-sensitivity marker of DNA double-strand breaks, in the urothelium of male F344 rats treated with N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN), a bladder-specific carcinogen. Our results clearly demonstrated that γ-H2AX aggregation was specifically generated in nuclei of bladder epithelial cells of BBN-treated rats, which was not found in untreated controls or mesenchymal cells. γ-H2AX-positive cells were detected not only in hyperplastic and neoplastic areas but also in the normal-like urothelium after BBN treatment. These data indicate that γ-H2AX has potential as a useful biomarker for early detection of genotoxicity in the rat urinary bladder. To the best of our knowledge, this is the first report demonstrating expression of γ-H2AX during bladder carcinogenesis.     

Associations among albuminuria, C-reactive protein concentrations, survival predictor index scores, and survival in 78 critically ill dogs

Background: Microalbuminuria and C‐reactive protein (CRP) are predictors of morbidity and survival in critically ill human patients. Hypothesis/Objectives: To evaluate results of microalbuminuria assays (untimed single‐sample urine albumin concentration [U‐ALB] and the urine albumin : creatinine ratio [UACR]), serum CRP, and survival predictor index (SPI2) scores as predictors of survival in critically ill dogs. Animals: Seventy‐eight dogs admitted to intensive care units at University of Tennessee (UT) and Colorado State University (CSU). Methods: Prospective observational study. Critically ill dogs were eligible for enrollment, unless euthanized because of financial constraints. Samples were collected within 3 hours of admission. Spearman's rank‐correlation coefficients were determined for U‐ALB, UACR, CRP, and SPI2. U‐ALB, UACR, CRP, and SPI2 were assessed for associations with 7‐ and 30‐day survival by Mann‐Whitney U‐tests and receiver operating characteristic (ROC) curves. P‐values < .0125 were considered significant. Results: UT (n = 49) and CSU (n = 29) patients did not differ significantly. Forty percent (31/78) of dogs died. SPI2 was inversely correlated with U‐ALB (r_s=?0.39, P < .001) and UACR (r_s=?0.41, P < .001). CRP was not correlated with SPI2 (P= .019), U‐ALB (P > .1), or UACR (P > .1). U‐ALB and UACR had very high correlation (r_s= 0.95, P < .001). SPI2, U‐ALB, and UACR differed significantly for survivors and nonsurvivors. SPI2, U‐ALB, and UACR had areas under the ROC curve (AUC) from 0.68 to 0.74 for survival prediction. Conclusions and Clinical Importance: Albuminuria and SPI2, but not CRP, are associated with survival in critically ill dogs. Suboptimal AUCs limit the value of microalbuminuria testing for clinical risk assessment. Additional studies are necessary to determine the usefulness of microalbuminuria testing in patient risk stratification for prospective research.     

人肝癌细胞系HepG2的次黄嘌呤鸟嘌呤磷酸核糖转移酶缺陷型的建立

本试验旨在建立次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)缺陷型的HepG2细胞系,为细胞杂交与单克隆抗体的研究提供亲本细胞。通过N-甲基-N-硝基-N-亚硝基胍(MNNG)诱导HepG2细胞突变,逐步提高培养液中6-巯基鸟嘌呤(6-TG)的浓度,筛选出对6-TG有抗性的细胞株,检测它在HAT中生长的情况。得到可在含20 μg/mL 6-TG的高糖DMEM完全培养基中稳定生长的细胞株,此细胞株在HAT培养基中第3天出现明显的死亡,第12天全部死亡,诱变后细胞的染色体分布于32-88,染色体众数为48。突变筛选出的细胞具有对6-TG抗性和对HAT敏感的特性,证实该细胞是HGPRT缺陷型细胞株,为以后进行杂交瘤制备提供了适宜的亲本细胞。     

不同CO2浓度对苜蓿分枝期光合性能的影响

以苜蓿（Medicago sativa）‘Maverick’为试验材料,研究不同CO₂浓度下苜蓿分枝期的光合能力日变化情况,探索未来大气CO₂浓度上升过程中苜蓿的生产潜力,为未来牧草种植产业提供指导。结果表明：当开顶式生长室中CO₂浓度为550和700 μmol·mol^-1时,‘Maverick’的叶绿素含量对比室外对照组分别提高了2.4%和3.6%,但差异不显著。净光合速率（Pn）日平均值分别提高了114.69%和116.54%。此外,CO₂浓度升高提高了苜蓿的光系统Ⅱ（PSII）的原初光能转换效率（Fv/Fm）、潜在活性（Fv/Fo）、实际光化学量子产量（Yield）,并显著增加了光化学淬灭（qP）,降低非光化学淬灭（NPQ）。研究结果表明在高CO₂浓度条件下更利于‘Maverick’叶绿素捕获光能,PSII反应中心比例大,光合作用增强。     

C型利钠肽及其受体在水牛卵泡中的表达分析

为了探明C型利钠肽（C-type natriuretic peptide,CNP）及其受体（natriuretic peptide receptor,NPR）在水牛卵泡中的表达模式,本研究首先采用实时荧光定量PCR技术检测水牛卵巢中利钠肽家族主要成员A型、B型、C型利钠肽（ANP、BNP、CNP）及其Ⅰ型、Ⅱ型受体（NPR1、NPR2）的mRNA表达情况,然后利用免疫组化技术对水牛卵泡中CNP及NPR2蛋白进行定位,最后利用实时荧光定量PCR技术检测颗粒细胞和卵丘细胞中CNP及NPR2的mRNA表达规律。结果显示,水牛卵巢主要表达CNP及NPR2,且在各级卵泡中均有表达,其中,CNP主要在壁层颗粒细胞中表达,NPR2主要在卵丘细胞中表达;颗粒细胞上CNP mRNA表达水平显著高于卵母细胞周围的卵丘细胞（P<0.05）,而卵丘细胞上NPR2 mRNA表达水平显著高于颗粒细胞（P<0.05）。综上所述,在利钠肽主要家族成员和受体中,CNP和NPR2在水牛卵巢中呈现强表达,CNP主要在壁层颗粒细胞中表达,而NPR2主要在卵母细胞周围的卵丘细胞中表达。     

保护性耕作对黄土旱塬玉米土壤呼吸及微生物数量的影响

研究了实施保护性耕作10年后秸秆还田和免耕对土壤可培养微生物数量和CO2释放的影响,采用LI COR 6400 09土壤气室连接红外线气体分析仪测定了玉米（Zea mays cv.Zhongdan No.2）土壤呼吸速率,采用平板涂抹计数法测定了根际土壤细菌、真菌和放线菌数量。结果表明,免耕和秸秆还田处理玉米花期土壤呼吸速率降低了8.3%和7.8%,玉米收获期土壤呼吸速率降低了15.5%和13.6%;土壤10 cm处地温与土壤呼吸速率变化趋势一致,且在传统耕作处理下最高;与传统耕作相比,免耕和秸秆还田不同程度地增加了根际土壤细菌、真菌和放线菌的数量,但各类微生物数量之间的比例恒定;土壤呼吸速率与3类根际土壤微生物的数量呈负相关,但不显著。可利用免耕和秸秆还田提高黄土旱塬土壤碳库贮量。     

Orf virus circulation in cattle in Turkey

Orf virus (ORFV) causes contagious skin disease that mainly affects sheep and goats with zoonotic potential. However, there is not enough information about the association between ORFV and occurrence of skin disease in cattle. The present study describes outbreaks of ORFV infection in cattle in different provinces that are located in the Aegean, Central Anatolian and Mediterranean regions of Turkey. During the months of June and August 2017, vesicular fluid and scab samples were collected from cattle which had proliferative skin lesions. First, presence of lumpy skin disease virus (LSDV) and bovine herpesvirus 2 (BoHV-2, known as the causative agent of pseudo-lumpy skin disease) were investigated by real time PCR and PCR, respectively. Then, samples tested for the presence of parapoxviruses by PCR using primers specific to major envelope protein gene (B2L). Parapoxvirus DNA was detected in investigated samples whereas LSDV and BoHV-2 DNA were not detected. The analysis of the B2L gene sequences revealed that cattle were infected with ORFV. The isolates in the present study shared 100% sequence identity at the nucleotide and amino acid level when compared with previously characterised Turkish field ORFV isolates from goats in 2016. Results of the study show unusual infection of cattle with ORFV, and suggest that ORFV jumps the host species barrier from goats to cattle.