Preliminary studies of serum acute-phase protein concentrations in hematologic and neoplastic diseases of the dog

Serum concentrations of acute-phase proteins (APPs): haptoglobin (Hp), ceruloplasmin (Cp), serum amyloid A (SAA), and C-reactive protein (CRP) were determined in healthy dogs (n = 15) and dogs with different diseases grouped as acute inflammation (I, n = 12), hematologic neoplasias (HT, including leukemia and lymphoma, n = 16), nonhematologic neoplasias (NHT, including epithelial, mesenchymal, and mixed, n = 20), and autoimmune hemolytic anemia (AIHA, n = 8). SAA and CRP were analyzed using commercially available enzyme-linked immunosorbent assay (ELISA) kits, and Hp and Cp were measured using colorimetric methods, all previously validated for use in dogs. Increased concentrations of all APPs were observed in all groups of diseased dogs, but statistical significance only was observed with Hp (I, P < .001; HT, P < .05), Cp (I, P < .05; AIHA, P < .01), and CRP (I, P < .001; HT, P < .001; AIHA, CRP P < .05). High variability in individual APPs within each group of diseases was found with no significant differences between leukemia and lymphoma as well as among different types of neoplasia. The AIHA group had smaller increases in Hp, SAA, and CRP but higher concentrations of Cp. When follow-up of individual cases was possible, a decrease in APPs generally was found in cases with favorable outcome. The results of this study suggest that neoplasia and hematologic diseases such as AIHA should be considered as possible causes of mild increases in APPs in dogs. Measurement of APPs may be helpful to assess clinical evolution and monitor treatment of these processes.     

Use of serum amyloid A in equine medicine and surgery

Serum amyloid A (SAA) has become an indispensable part of the management of equine patients in general practice and specialized hospital settings. Although several proteins possess acute phase properties in horses, the usefulness of SAA exceeds that of other acute phase proteins. This is due to the highly desirable kinetics of the equine SAA response. SAA concentrations exhibit a rapid and pronounced increase in response to inflammation and a rapid decline after the resolution of inflammation. This facilitates the detection of inflammatory disease and real-time monitoring of inflammatory activity. SAA may be used in all stages of patient management: (1) before diagnosis (to rule in/rule out inflammatory disease), (2) at the time of diagnosis (to assess the severity of inflammation and assist in prognostication), and (3) after diagnosis (to monitor changes in inflammatory activity in response to therapy, with relapse of disease, or with infectious/inflammatory complications). By assessing other acute phase reactants in addition to SAA, clinicians can succinctly stage inflammation. White blood cell counts and serum iron concentration change within hours of an inflammatory insult, SAA within a day, and fibrinogen within 2–3 days; the interrelationship of these markers thus indicates the duration and activity of the inflammatory condition. Much research on the equine SAA response and clinical use has been conducted in the last decade. This is the prerequisite for the evidence-based use of this analyte. However, still today, most published studies involve a fairly low number of horses. To obtain solid evidence for use of SAA, future studies should be designed with larger sample sizes.     

D-Dimer Improves the Prognostic Value of Combined Clinical and Laboratory Data in Equine Gastrointestinal Colic

The discriminating ability of 15 parameters alone or in combinations, including results from analysis of plasma endotoxin, the Nycomed plasma D-Dimer test and phospholipase A₂, were analyzed to predict morbidity and mortality in equine gastrointestinal colic. Endotoxaemia was a characteristic feature of the colic horses. The problem of adequately predicting non-survivors among colic horses required several parameters to be included in the logistic model: if the “classical parameters”, (heart rate, respiratory rate, PCV, anion gap) were included in the model, addition of plasma D-dimer, phospholipase A₂, and Cl^- significantly improved the predictive value of the logistic model. Increasing heart rate and D-dimer together with decreasing chloride was a risk factor for nonsurvival. The sensitivity of this three-parameter logistic model to predict nonsurvival was 78% and specificity 77%. The Nycomed D-Dimer test is recommended as a horse-site test to predict disseminated intravascular coagulation and nonsurvival in equine colic.     

Exogenous testosterone modulates tumor necrosis factor-alpha and acute phase protein responses to repeated endotoxin challenge in steers

Clinical responses to some disease agents differ between sexes and this dimorphism has been attributed to the immunomodulating effects of steroid hormones. Our objective was to determine in steers the effect of testosterone on circulating concentrations of immune response mediators (tumor necrosis factor-alpha, TNF-alpha; serum amyloid-A, SAA; haptoglobin, HG; xanthine oxidase, XO; nitric oxide, NO) after two consecutive endotoxin challenges (LPS1 and LPS2, 5 days apart; 0.25 microg/kg BW). Sixteen crossbred steers (328+/-6 kg) were assigned to control (CON, n=8) or testosterone cypionate treatment (TES, n=8; 100 mg/m2 body surface; i.m. injection 12 and 2 days before LPS1). The response to LPS was calculated as area under the timexconcentration curve (AUC) for the parameter measured. After LPS1, TNF-alpha AUC was greater in TES than CON (P<0.05). Plasma HG and SAA concentrations increased (P<0.01) after LPS1 and LPS2. In all steers SAA AUC was greater after LPS1 than LPS2 (P<0.01) but the response was augmented over CON with testosterone treatment (P<0.05). HG response to LPS1 within 24 h was not affected by testosterone. However, 5 days after LPS1 mean plasma HG concentration remained higher in TES than CON (P<0.01). HG response to LPS2 was greater in TES than CON (P<0.01). Plasma nitrate+nitrite concentration (NO production marker) and XO activity increased after each LPS challenge but responses were not affected by testosterone treatment. Results indicate that the presence of circulating testosterone increases the magnitude of the TNF-alpha response to LPS challenge as well as the subsequent increases in acute phase proteins (APP). Effects of testosterone on increases in TNF-alpha and APP may underlie a differential presentation of disease symptoms between sexes or between steers and bulls. The data also suggest a role for testosterone in the development of tolerance to repeated immune challenge through its effect on the increased magnitude and duration of HG response.     

Paraoxonase activity as a tool for clinical monitoring of dogs treated for canine leishmaniasis

This study was designed to determine if the activity of paraoxonase (PON1), an antioxidant enzyme that works as a negative acute phase reactant, is a better predictor for the clinical recovery of leishmaniotic dogs receiving standard treatments compared with inflammatory markers such as C reactive protein (CRP) and electrophoretic fractions. For this purpose we tested 20 healthy dogs (controls) and 39 leishmaniotic dogs classified as sick (group A, n = 23) or severely sick (group B, n = 16) and tested at admission and after 3, 7, 14, 21, 28, 35 and 42 days.At admission, CRP and electrophoresis were altered in both groups, while PON1 activity was abnormal only in group B. There were no differences related to the outcome (mortality, complications or time of recovery). PON1 activity normalized in about 2 weeks in dogs that had abnormal values at admission and a final positive outcome; CRP normalized in 4–6 weeks and electrophoretic fractions were still altered after 6 weeks. The results show that, at admission, inflammatory markers did not predict the outcome of leishmaniasis. PON1 activity decreased only in some dogs with systemic inflammation but not in those with mild leishmaniasis: when decreased, PON1 normalized earlier than other markers in dogs that responded to treatment. This finding most likely depends on the rapid decrease in oxidative phenomena. PON1 activity should therefore be tested on admission: if low values are recorded, severe inflammation may be suspected and PON1 measurement may be repeated during treatment to early identify responsive dogs.     

促凋亡基因(Bad)在卵泡期绵羊生殖器官中的表达

以卵泡期小尾寒羊和萨福克羊为研究对象,采用免疫组织化学技术,对促凋亡基因(Bad)在生殖器官卵巢、输卵管和子宫中的表达与定位进行初步研究。结果表明,在卵巢组织中Bad基因在原始卵泡、初级卵泡和次级卵泡中呈中度着染,在三级卵泡中呈轻度着染,阳性细胞在卵泡上皮细胞和颗粒细胞中分散分布,呈轻度着染;输卵管上皮分泌细胞和部分纤毛细胞胞质中见Bad基因中等强度表达,输卵管壶腹部上皮中,Bad基因多表达于分泌细胞,纤毛细胞中有部分呈阳性且强度弱,宫管结合部、峡部和壶腹部Bad基因阳性细胞染色强度弱,均呈中度着染;子宫内膜子叶和基质中的阳性细胞数量极少,阳性细胞呈轻度着色。利用计算机图像分析技术测量小尾寒羊和萨福克羊生殖器官各组织细胞中Bad基因表达的阳性细胞率和平均光密度,结果Bad基因在这2品种绵羊组织中的表达规律大体相似,但也存在一些明显的差异:萨福克羊三级卵泡颗粒细胞Bad基因阳性细胞率显著高于小尾寒羊(P0.01);萨福克羊三级卵泡和成熟卵泡膜细胞Bad基因阳性细胞率和平均光密度均显著高于小尾寒羊(P0.01);萨福克羊子宫内膜腺体卵泡期Bad基因阳性细胞平均光密度显著高于小尾寒羊(P0.01);由此表明,小尾寒羊和萨福克羊卵泡期生殖器官中细胞凋亡的差异与Bad基因表达的差异,可能是造成两者繁殖力高低差异的基础。     

伊维菌素纳米乳透皮制剂的研究

本试验研制伊维菌素纳米乳透皮制剂,并对其理化性能、稳定性、体外药物释放及透皮性能进行评价。采用三元相图筛选不同载药量的纳米乳,确定最佳载药量;采用响应面优化设计筛选纳米乳处方,考察了伊维菌素纳米乳的平均粒径、电位、形态、pH、黏度等性能;分别采用透析袋法和Franz扩散池法比较伊维菌素纳米乳透皮制剂与市售伊维菌素皮肤涂剂的体外释放行为和透皮性能。结果显示,载药量为2.00%时,纳米乳区域最大、最稳定;获得的优选处方为聚氧乙烯蓖麻油 :二乙二醇单乙基醚 :油酸乙酯 :伊维菌素 :水=26 :12 :7: 2: 53;所得伊维菌素纳米乳的平均粒径为18 nm;伊维菌素纳米乳在室温条件和4 ℃冰箱中保存1年仍稳定;其24 h皮肤累积渗透量和滞留量分别是市售伊维菌素皮肤涂剂的3.24和2.05倍。研究结果表明,研制的新型伊维菌素纳米乳透皮制剂具有制备工艺简便、稳定性好、透皮性能好等优点,具有很好的应用前景。     

中国草地资源经营的历史发展与当前任务

回顾近40年中国草地资源经营发展历程,分析了存在问题及其原因,提出了当前任务:1.要明确工作总则,围绕生态与小康两大目标,思考与部署草地资源经营工作;确立草地生态置换理论与对策;组建草原生态建设与草原牧业发展综合治理工程.2.要做好退牧还草工程,主要是把握实质做出实效;做好统筹规划、加强综合治理;因地制宜,做好实施方案;提高科技含量,做好基础工作,特别是治理区水、草、林、土资源本底重估与优化配置,草牧产业发展要点,以及牧民定居点建设工作.     

C18-磁性纳米颗粒萃取虾肉中氯霉素残留

建立了一种用C18键合磁珠固相萃取虾肉中氯霉素残留的方法。在Fe3O4超顺磁性纳米颗粒的表面键合C18基团,制备双功能反相萃取颗粒。用该颗粒对样品中的氯霉素残留进行固相萃取,对结合时间、温度、磁珠用量等因素进行了优化,建立磁珠法兽药残留固相萃取方法。用标准方法(SN/T 1864-2007)对萃取所得产物进行检测。经过优化,C18键合磁珠的最佳萃取条件为50℃结合5 min,用1 mL甲醇洗脱3次。检测结果显示该方法具有良好的精密度和回收率。在0.1～10μg/kg之间具有良好的线性关系,相关系数为0.9941,检出限为0.1μg/kg。该方法灵敏度高、重现性好、准确度高,可满足虾肉中氯霉素残留检测的需要。