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.     

猪圆环病毒2型IgM抗体间接ELISA的建立

猪圆环病毒2型(PCV2)是断奶仔猪多系统衰竭综合症的重要原发性病原,其感染的早期检测有助于及时采取防控措施。以重组衣壳蛋白(Cap蛋白)作为包被抗原,通过对反应条件的优化,建立了PCV2 IgM抗体间接ELISA,其最适抗原包被浓度为1.25 μg／mL,最适血清稀释度为1∶100,临界值为0.35。该检测方法与其他5种常见猪疫病阳性血清无交叉反应,特异性良好。批内、批间重复试验的变异系数均在2%~6%,重复性好。对100份临床样本的检测结果表明,该检测方法与国外同类试剂盒相比,敏感性为90.3%,特异性为92.8%,总符合率为92%。试验结果表明,所建立的PCV2 IgM间接ELISA特异性好和敏感性高,适用于PCV2感染早期的流行病学调查。     

融合表达猪圆环病毒2型ORF1和ORF2基因真核质粒的构建及对小鼠的免疫原性

用PCR法扩增出猪圆环病毒2型的Rep蛋白基因（933bp）和Cap蛋白基因（705bp）,将其定向克隆于真核表达载体pcDNA3.1（＋）的多克隆位点上,构建真核表达质粒pcDNA-ORF1-ORF2。将构建好的重组质粒pcD-NA-ORF1-ORF2按100μg/只腿部肌肉注射BALB/c小白鼠,同时设pcDNA-ORF1、pcDNA-ORF2、pcDNA3.1-（＋）、PCV2全毒疫苗和PBS为对照,共免疫2次,间隔2周。分别于首免后第0、7、14、21、28、42天用MTT法检测小鼠脾淋巴细胞的增殖效应,用ELISA法检测小鼠的抗体水平;并于首免后第0、7、14、21、28天测定脾淋巴细胞中各细胞亚群的比例,对该核酸疫苗的免疫原性进行初步评价。结果显示,重组质粒能诱导鼠体产生较强的细胞免疫和体液免疫,并从免疫后第7天起所测各组数据均显著高于（P〈0.05）或极显著高于（P〈0.01）其他试验组。结果表明,将ORF1和ORF2基因共同用于PCV2核酸疫苗的研发具有很好的前景,为研究新型猪圆环病毒疫苗奠定了基础。     

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。     

华北山地阳坡中生灌草植被对CO2浓度和温度变化的光合响应

利用便携式光合气体分析系统(LI-6400),从全球变化的角度分析华北山地阳坡灌草植被群落中5种具有代表性的植物光合特征对CO2浓度和温度变化的响应.结果表明:随着温度的升高,狗尾草Setaria viridis和龙牙草Agrimonia pilosa净光合速率呈明显增大趋势,在华北山地阳坡灌草群落中具有较强的竞争力,而达乌里胡枝子Lespedeza davurica的净光合速率对CO2的响应值降低,在环境温度升高的条件下不利于其光合产物的积累.     

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.     

HSV-2gD融合gB CTL表位重组蛋白的原核表达及免疫原性评价

为研制具有较强细胞免疫作用的单纯疱疹病毒Ⅱ型(HSV-2)疫苗,将HSV-2糖蛋白D(gD)序列与糖蛋白B(gB)上两种CTL表位序列连接,人工合成重组基因gD-gBCTL,通过PCR扩增,将其克隆后连接至原核表达栽体pET-22b(+),构建重组基因表达质粒pET-22b(+)-gD-gBCTL,转化E.coli B...