3种禽血抗凝剂效果及其对DNA提取质量的影响

通过3种不同血液抗凝剂对禽血抗凝,提取新鲜的和经冷冻保存一段时间后的抗凝血DNA,运用紫外分光法、凝胶电泳和PCR扩增技术检测提取的DNA样品质量,探讨不同抗凝剂的抗凝效果及其对提取DNA质量的影响。结果表明:3种抗凝血提取的DNA纯度和浓度相近。新鲜抗凝血比冷冻保存一段时间后的抗凝血提取的DNA质量好。ACD抗凝血在冷冻一段时间后出现凝血块,EDTA抗凝血液放置时间过长也影响抗凝效果,但两者在加入裂解液后则可长时间保存。75%乙醇抗凝剂抗凝效果较好,可长时间保存,适合远距离采血。由3种抗凝禽血提取的DNA分子片段完整,能满足常规分子生物学试验研究。     

Pulmonary Thromboembolism in 29 Dogs: 1985–1995

Pulmonary thromboembolism (PTE) occurs as a complication to a number of commonly encountered clinical diseases. Antemortem recognition of this life-threatening disorder is hampered by nonspecificity of clinical signs. This retrospective study was performed to analyze clinical features, laboratory findings, imaging abnormalities, and concurrent postmortem diagnoses in 29 dogs with confirmed pulmonary embolism. A variety of clinicopathologic and radiographic abnormalities were noted but there were no pathognomonic findings for PTE. Arterial blood gas analyses were performed in 15 (52%) of 29 dogs; 12 (80%) of 15 exhibited hypoxemia and 15 (100%) of 15 had increased alveolar-arterial oxygen gradients. Response to supplemental O2 was variable and did not correlate with the presence or absence of additional pulmonary pathology on postmortem. At postmortem, 25 (86%) of 29 dogs had grossly visible emboli, 17 (59%) of 29 dogs had multiple disease processes, and 16 (55%) of 29 dogs had additional pulmonary pathology. PTE was suspected antemortem in 11 (38%) of 29 dogs. In dogs with respiratory signs consistent with PTE, the condition was a differential diagnosis in 11 of 17 animals; all had diseases previously reported to be associated with PTE. Neoplasia, systemic bacterial disease, and immune-mediated hemolytic anemia were diagnosed most frequently.     

Plasma biochemistry of ostrich (<Emphasis Type="Italic">Struthio camelus</Emphasis>): effects of anticoagulants and comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals, but limited information is existing for ostrich plasma biochemistry. Ten clinically healthy ostrich were blood sampled in different tubes containing each anticoagulant and plain tube for harvesting plasma and serum. The concentrations of glucose, cholesterol, uric acid, creatinine, total protein, albumin, calcium, inorganic phosphorus, and magnesium and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma glutamyl transferase (GGT) were measured. The concentrations of glucose, uric acid, total protein, and calcium were significantly lower in citrated plasma than that of serum. For dilution corrected citrated plasma significant differences were only seen for the concentration of uric acid. Most parameters did not show any differences, but significant increase were seen for glucose, total protein, albumin, and phosphorus concentrations when heparin was used as an anticoagulant.     

3种抗凝剂对大菱鲆血液抗凝效果、血细胞形态和血液生理生化指标的影响

鱼类血液极易凝结,经抗凝剂处理后的血液可充分满足各项生理指标检测,为衡量鱼体各项生理机能提供重要参考,同时,鱼类种属多样,在抗凝剂的选择和使用上存在一定差异。基于此,本研究以欧亚养殖良种大菱鲆(Scophthalmus maximus)为研究对象,选用乙二胺四乙酸二钾、肝素钠和柠檬酸钠3种抗凝剂,通过对大菱鲆正常状态和急性低氧胁迫条件下[溶解氧浓度为(1.2±0.3) mg/L]抗凝效果评价、血细胞形态观察和血液生理生化指标分析,筛选急性低氧胁迫条件下最佳血液抗凝剂。结果显示,正常溶解氧状态下,乙二胺四乙酸二钾抗凝效果显著,急性低氧胁迫下静置6 h和12 h后,抗凝效果显著抗凝剂分别为乙二胺四乙酸二钾和肝素钠。正常溶解氧状态肝素钠抗凝剂下血细胞出现双核现象,柠檬酸钠和乙二胺四乙酸二钾抗凝剂下分别出现嗜酸性、嗜碱性粒细胞和嗜碱性粒细胞,急性低氧胁迫导致乙二胺四乙酸二钾、肝素钠、柠檬酸钠抗凝剂均观察到双核血细胞,其中,肝素钠和柠檬酸钠抗凝剂还分别出现微核和无核血细胞。急性低氧胁迫处理后,3种抗凝剂作用下血液白细胞数目均显著增高(P<0.05),乙二胺四乙酸二钾和肝素钠抗凝作用下,红细胞数目和血红蛋白含量显著升高(P<0.05),而柠檬酸钠抗凝剂作用下,红细胞数目和血红蛋白含量显著降低(P<0.05)。3种抗凝剂作用下血浆葡萄糖和皮质醇浓度显著增高(P<0.05),但肝素钠抗凝剂作用下,葡萄糖和皮质醇浓度显著低于柠檬酸钠和乙二胺四乙酸二钾(P<0.05)。综上所述,乙二胺四乙酸二钾可作为急性低氧胁迫条件下大菱鲆血液学分析的首选抗凝剂。     

The influence of anticoagulants on the measurement of total protein concentration in equine peritoneal fluid

The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The solution of K3EDTA in distilled water only caused TP overestimation at high K3EDTA concentrations (>5 micromol/ml). By contrast, lithium heparin did not influence the refractometric values of TP. Neither anticoagulant modified TP values when measured by the biuret method. In conclusion, the use of K3EDTA as anticoagulant may result in a significant overestimation of TP values of peritoneal fluid samples measured by refractometry.     

Identification of cytochrome P450 differentiated expression related to developmental stages in bromadiolone resistance in rats (Rattus norvegicus)

Adult, 20-week-old, rats from a Danish bromadiolone-resistant strain of rats (Rattus norvegicus) over-express the cytochrome P450 genes Cyp2e1, Cyp3a2 and Cyp3a3 upon bromadiolone exposure. Furthermore, adult female rats of this strain over-express the Cyp2c13 gene and suppress Cyp2c12, while males over-express the Cyp2a1 gene. The altered gene expression has been suggested to be involved in the bromadiolone resistance by facilitating enhanced anticoagulant metabolism. To investigate the gene expression of these cytochrome P450 genes in rats of different developmental stages we compared expression profiles from 8-, 12- and 20-week-old resistant rats of the Danish strain to profiles of anticoagulant-susceptible rats of same ages. The three age-groups were selected to represent a group of pre-pubertal, pubertal and adult rats. We found expression profiles of the pre-pubertal and pubertal resistant rats to concur with profiles of the adults suggesting that cytochrome P450 enzymes are involved in the Danish bromadiolone resistance regardless of developmental stage. We also investigated the relative importance of the six cytochrome P450s in the different development stages of the resistant rats. The P450-3a2 and -3a3 isoforms were proposed to be of higher importance in adult male resistance than in pre-pubertal resistance. In contrast, the P450-2c13 and -3a2 isoforms were proposed to be more important in sexual immature female resistance, while the P450-2e1 and -3a3 isoforms were suggested to play a more significant role in adult female resistance.     

Effects of heparin, citrate, and EDTA on plasma biochemistry of sheep: Comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals, but limited information is existing for sheep plasma biochemistry. Ten clinically healthy Baloochi breed of sheep were blood sampled in different tubes containing each anticoagulants and plain tube for harvesting plasma and serum. The concentrations of glucose, cholesterol, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, and magnesium and the activity of aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of GGT, bilirubin and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than that of serum. For corrected citrated plasma significant differences were seen for the concentrations of glucose, creatinine, calcium and the activity of ALP.Most parameters did not show any difference, but significant increase was seen for albumin concentration when heparin was used as an anticoagulant. Using EDTA as anticoagulant caused a significant difference for the concentrations of some of the measured parameters in plasma except glucose, GGT, cholesterol, albumin, bilirubin, CK, and inorganic phosphorus comparing with serum.