Enhancement of phagocytosis in bovine milk leukocytes in vitro

The data presented show a highly significant increase in numbers of bovine milk leukocytes, obtained from mammary glands of 13 cows irritated by sterile distilled water, which ingested staphylococci in four hr when 0.04% glucose was added to the leukocyte: staphylococci mixture. This increase was of the same order as that resulting from the addition of 5% of bovine anti-staphylococcal serum containing 16 I U of anti-alpha haemolysin. Nonimmune serum produced no enhancement. Cells from two other cows did not respond to either glucose or immune serum.     

Phagocytosis and related phenomena in polymorphonuclear leukocytes from cow's milk

There were fewer efficient phagocytes among leukocytes collected from artificially irritated mammary glands than among the leukocytes from blood of the same animals. The milk polymorphonuclear (PMN) leukocytes adhered poorly to a column of siliconised glass beads when compared with the blood cells. However, investigations of the O(2) uptake and CO(2) production of the milk PMN leukocytes revealed that these cells appeared to utilize metabolic pathways similar to those used by human and guinea pig PMN leukocytes during phagocytosis. These pathways are associated with degranulation and the production of H(2)O(2) following particle ingestion. It is therefore suggested that the milk PMN leukocytes appear not to have lost the ability to produce this bactericidal substance.     

Separation of mononuclear leukocytes and polymorphonuclear leukocytes from equine blood.

The present study describes a two step technique for the separation of mononuclear leukocytes or mononuclear and polymorphonuclear leukocytes from whole equine blood. First, the leukocyte rich plasma was obtained by sedimentation of erythrocytes in the undiluted blood. Subsequently, separation of the different populations of white blood cells was performed by centrifugation with different gradients overlaid with the leukocyte rich plasma. The optimal separation of the mononuclear cells was obtained by the centrifugation of the leukocyte rich plasma overlaying the gradient containing 24 parts of 9.5% ficoll and ten parts of 34% isopaque. The mononuclear leukocytes (95% lymphocytes and 5% monocytes) formed a monolayer band at the plasma-ficoll-isopaque interface and other blood cells migrated to the bottom of the tube. For the separation of mononuclear and granular leukocytes from the blood, the gradient containing 24 parts of 10% ficoll and ten parts of 34% isopaque was used. The separated monuclear leukocytes responded to stimulation with phytohemagglutin and viability of both mononuclear and polymorphonuclear leukocytes was not affected by ficoll-isopaque separation.     

牛奶和奶粉中八种镇定剂残留量的液相色谱-串联质谱测定

采用液相色谱-串联质谱仪,建立了牛奶和奶粉中乙酰丙嗪cetopromaizine、氯丙嗪chlor-promazin、氟哌啶醇haloperidol、丙酰二甲氨基丙吩噻嗪propionylpromazine、甲苯噻嗪xylazine、阿扎哌隆azaperone及其代谢物阿扎哌醇azaperol和咔唑心安carazolol残留量的液相色谱-串联质谱测定方法。结果表明八种镇定剂在牛奶中添加水平0.10～0.50μg/kg到0.80～4.00μg/kg,其回收率在86.29%～92.49%之间,相对标准偏差在8.99%～20.56%之间;在奶粉中添加水平0.80～2.00μg/kg到2.0～8.00μg/kg,其回收率在81.59%～97.49%之间,相对标准偏差在1.76%～16.70%之间。八种镇定剂在确定的添加范围内线性关系良好。八种镇定剂在牛奶中的方法检出限为0.1～0.5μg/kg,在奶粉中的方法检出限为0.8～4.0μg/kg。     

Phagocytic acitivity of bovine leukocytes during pregnancy.

The phagocytic competence, measured as the total number of polymorphonuclear leukocytes per mm3 which phagocytosed Staphylococcus aureus, strain 321, in vitro, was determined in eight cows during complete pregnancies. Such leukocytes are referred to as "Active PMN'S". There was a gradual decline in the number of these cells from conception to a minimum between the 16th and 20th weeks of pregnancy, followed by a steady increase to the cessation of lactation when a marked drop occurred, after which there was an increase to a maximun during the second week prepartum. From this maximum there was a rapid decrease to an absolute minimum during the first week after parturition. From the second week postpartum there was a gradual increase to conception. The correlation coefficient (r) of number of active PMN'S with time before conception was -0.474 )p-0.01). There were significant differences (p=0.01) in numbers of active PMNS Among the eight cows. It was found that the cows fell into two groups, one whose members had, overall, significantly more active PMNs (p=0.001) than those in the second group. The between cow differences may have been due to 1) age, since the cows with the highest numbers of circulating active PMNs were younger than those in the other group of 2) the combined stress of pregnancy and lactation, as those cows which were both pregnant and milking had the lowest numbers of active PMNs.     

肺癌患者外周血白细胞端粒DNA相对长度检测

目的:探讨外周血白细胞端粒DNA相对长度与肺癌的关系。方法:采用SYBRGreenⅠRealTimePCR方法检测54例肺癌患者(病例组)和77例健康对照(对照组)外周血白细胞端粒DNA的相对长度。结果:病例组外周血白细胞端粒DNA的相对长度为(3.43±0.87),短于对照组(3.86±1.46),差异有统计学意义(t=2.093,P=0.038)。进一步应用logistic回归分析,把端粒长度按百分位数P33和P66从短到长依次分为T1、T2、T3三层,调整吸烟与年龄因素后,以T3层为参考水平,结果显示T2层患肺癌危险度为3.933(95%CI1.139～13.580),T1层患肺癌危险度为4.684(95%CI1.377～15.936),P<0.05。结论:端粒DNA相对长度缩短可能与肺癌的发生有关。     

Mexiletine in human blood and breast milk.

Twelve paired samples of breast milk and blood were obtained, between the 2nd and 5th days after she had given birth, from a 30-year-old woman who was a 8-hourly propranolol (20 mg) and mexiletine (200 mg). Propranolol was detected in 9 of the samples of blood and in only 4 of the milk samples. Mexiletine was measurable in all samples, and the milk : plasma ratio varied between 0.79 and 1.89 with a mean of 1.45. However, the large volume of distribution of mexiletine makes it unlikely that the small dose of the drug received from the milk would be detrimental to the health of infants.     

The effect of added serum and glucose, and some inherent factors, on phagocytosis in vitro by milk leukocytes from several cows

On four occasions leukocytes were obtained from mammary secretions of four cows following irritation by sterile distilled water.The effects of three levels of bovine immune serum, three levels of glucose, and combinations of these on the ability of the leukocytes to ingest staphylococci in vitro were studied. Phagocytosis was estimated by examination of stained slides prepared after four hours contact at 37 degrees C.While 1% immune serum increased the percentage of milk leukocytes ingesting staphylococci, no improvement resulted from the addition of 3% or 5%. Addition of 20mg% glucose produced a larger increase than did 1% serum, and 40 and 60mg% gave further increases. Little advantage was obtained by combining serum and glucose at any of the levels studied.Overall significant differences were found in the ability to ingest staphylococci by milk leukocytes from the four cows, and the previous observation that cells from some cows in vitro do not respond fully to added serum and glucose was confirmed. Neither of these phenomena was related to glycogen levels in blood or milk leukocytes as determined by the anthrone method, blood glucose levels, leukocyte viability, relative proportions of PMN's in circulating blood white cells, nor the amount of homologous blood serum in the mammary secretions. In two cows whose milk cells did not respond fully, very few blood PMN's were active.     

Comparison of some storage and isolation methods to recover bluetongue virus from bovine blood

Bovine blood containing bluetongue virus was stored as whole blood with EDTA (4 degrees C), as washed cellular components (4 degrees C), and as washed cellular components with 10% DMSO (-70 degrees C). Periodic isolation attempts were made over a period of 330 days in four cell lines and embryonating chicken eggs (intravenous inoculation). Bluetongue virus was successfully isolated in all systems from most samples throughout the test period. There appeared to be more variation amongst days of attempted isolation within systems than between systems.     

Homocysteine induces expressions of adhesive molecules on leukocytes in whole blood

Objective To observe whether homocysteine can directly alter the expressions of CD11b, CD1 8, CD14 and L-selectin on neutrophils, monocytes, and lymphocytes in whole bloo d from healthy human subjects. Methods Leukocyte surface adhesive molecule expressions were analyzed by immunofluoresce nce flow cytometry.Results Homocysteine at the lowest concentration (20 μmol/L) significantly increased s urface adhesive molecule expressions of CD11b and CD18 on each cell type and CD1 4 on monocytes and neutrophils in whole blood.These effects were increased at homocysteine concentrations of 200 and 400 μmol/L, but at concentrations 1 mmol/L, CD11b/CD18 and CD14 expressions on all types of leukocytes were decrea sed.L-selectin expression was slightly decreased on all cell types in whole blood by homocysteine.Conclusion Homocysteine alters leukocyte expressions of CD11b/CD18, CD14 and L-selectin on leukocytes, which may be involved into homocysteine-induced leukocyte adhesion and migration.     

传染性非典型肺炎病人外周血白细胞损伤的病理学研究

目的　研究传染性非典型肺炎严重急性呼吸综合征 (SARS)病人外周血中白细胞的病理变化 ,及与SARS冠状病毒感染的关系 ,为探讨免疫细胞损伤在SARS发病机制中的作用提供形态学的依据。方法　采用 2 2例初步诊断的SARS病人外周血标本 ,及 4例正常成人外周血标本 ,分离收集所有的白细胞 ,进行透射电镜、免疫电镜和RT PCR检查。结果　白细胞的数量及亚型比例改变 :每个病例观察 10 0个细胞 ,分别计数粒细胞、淋巴细胞和单核细胞的数目。与正常组对照 ,粒细胞数目增多 (P <0 .0 5 ) ,淋巴细胞明显减少 (P <0 .0 5 ) ,T淋巴细胞 (CD4 5RO阳性 )仅占残存白细胞总数的 6 %～ 7%。在外周血白细胞胞浆内发现冠状病毒样颗粒者 6例 ,与RT PCR阳性者重合比例为5∶6。分别计数在各类白细胞中可观察到冠状病毒样颗粒的阳性细胞数 ,发现淋巴细胞中可观察到病毒的阳性细胞数最高。SARS病人血白细胞中的冠状病毒样颗粒为圆形 ,或略呈卵圆形 ,80～ 12 0nm左右 ,致密的双层质膜包绕 ,核心较疏松 ,膜外有放射状排列的纤细突起。散在分布于胞浆或粗面内质网、高尔基器及膜包绕的空泡中。淋巴细胞有变性和活化增殖的现象。结论　在SARS病人的血白细胞 ,内观察到冠状病毒样颗粒。提示 ,血细胞特别是淋巴细胞 ,也可能是冠状病毒攻     

同型半胱氨酸增高血白细胞表面粘附分子的表达

目的　观察同型半胱氨酸是否改变体外健康人血单核细胞、中性粒细胞及淋巴细胞表面粘附分子CD11b ,CD18,CD14和L 选择素 (L selectin)的表达。方法　细胞表面粘附分子CD11b ,CD18,CD14和L selectin的测定采用直接免疫荧光染色 ,2 4小时内用流式细胞仪测定。结果　同型半胱氨酸在低浓度 (2 0 μmol/L)时明显增加CD11b和CD18在各种白细胞表面的表达 ,同时也增加了CD14在单核及中性粒细胞的表达。这种作用随同型半胱氨酸浓度升高而增强 ,但当浓度升至≥ 1mmol/L时 ,各种粘附分子的表达反而降低。同型半胱氨酸使各种细胞类型表面L selectin降低。结论　同型半胱氨酸可改变白细胞表面粘附分子的表达 ,可能在体内通过此途经导致白细胞粘附并游出于血管内皮。     

血糖、白细胞和C-反应蛋白水平在急性有机磷中毒患者静脉血中的变化

目的：探讨急性有机磷农药中毒患血糖，白细胞，中性粒细胞和C－反应蛋白水平的变化及临床意义。方法：患入抢救室后，根据中毒的程度分为轻度，中度，重度3组，健康体检作为对照组，分别抽静脉血检测血糖，白细胞，中性粒细胞和C－反应蛋白。结果：中度和重度中毒组血糖，白细胞，中性粒细胞和C－反应蛋白水平均显高于对照组（P＜0．01，P＜0．05），轻度中毒组各项指标无显差异。重度中毒组随病情加重，各项指标呈上升趋势，中度中毒组则随病情减轻，各项指标呈下降趋势。结论：急性有机磷农药中毒患血糖，白细胞，中性粒细胞和C－反应蛋白水平的升高与中毒的程度呈正相关，血糖，白细胞，中性粒细胞和C－反应蛋白的水平越高，并发症及病死率也越高。这对指导治疗，减少并发症的降低病死率有一定意义。     

脂蛋白脂肪酶单克隆抗体的制备及特征分析

用牛奶纯化的脂蛋白脂肪酶(LPL),一分子量为56KD,免疫Balb/c小鼠,利用杂交瘤技术,建立了30余株分泌抗朱LPL单克隆抗体的杂交瘤细胞。对其中2E_5、2G_(10)、6D_7、8D_2、7F_4进行特征分析表明:①其抗体类别均为IgG,亚类分别为IgG_1、TgG_(2b)、IgG_(2b)、IgG_1、IgG_1;②抗体效价(细胞培养上清)5×10~(-2)——2×10~(-3);③5种单抗可认为识别三个不同的抗原表位;④Western—blot及Dot—blot结果显示各单抗均能识别纯化的LPL,与人奶均无交叉反应,与牛奶显示不同的结合特性。     

Occurrence of blood cells and serum proteins in bovine fetuses and calves

Hematological values of peripheral blood were determined for bovine fetuses and calves of various ages. Erythrocyte values increased through gestation. Fetuses 100 days or older had total values within the ranges of those reported for normal adult cattle. Mature erythrocytes were not observed in embryos and only a few were observed in fetuses 40 days of age. Fetuses 250 days or older had only a few rubricytes (<10/100 WBC). Leukocytes were first identified in the peripheral blood of a 45-day old fetus. Absolute leukocyte values increased through gestation and reached maximum values shortly before parturition. Granulocytes were first observed at 130 days of gestation and reached maximum values near parturition.Total serum protein and gamma-globulin concentrations of colostrum-deprived calves were similar to serum protein and gammaglobulin concentrations of fetuses older than 265 days and were lower than values for the colostrum-fed calves. The immunoelectrophoretic pattern of 59-day old fetuses, the earliest age at which serum samples were obtained, demonstrated albumin, an alpha1 globulin and a beta globulin, possibly transferrin. Additional alpha and beta globulins appeared in the older fetuses and by 175 days of gestation serum electophoretic patterns of the fetuses were similar to patterns normally found with adult bovine serum except for the absence of the gammaglobulins in fetal serum. Immunoglobulin M was detected in 39 of 95 fetal serum samples by radial diffusion and in 13 of 95 samples by immunoelectrophoresis. Immunoglobulin G was detected in ten of 95 fetal serum samples by radial diffusion and in six of 95 samples by immunoelectrophoresis.     

Somatic cell counts in bovine milk: relationships to production and clinical episodes of mastitis

The relationships between somatic cell counts, milk production and episodes of clinical mastitis were evaluated using data collected between 1979 and 1981 in 32 southern Ontario Holstein herds. Somatic cell counts were logarithmically transformed and the distribution of the resulting counts is presented. The seasonal pattern in cell counts was evaluated using a formal statistical procedure. Counts were lowest in the winter and spring and highest in the early fall but the differences amongst monthly geometric mean cell counts were small. Assuming a linear relationship between log somatic cell counts and test day milk production it was found that a unit increase in the log count resulted in a loss of 1.44 kg of milk. Regression analyses within specific log cell count ranges indicated that the previous estimate may underestimate losses at low cell counts and overestimate losses at higher cell counts. The relationships between cell counts and episodes of mild or acute clinical mastitis were evaluated by comparing counts preceding and following the clinical episodes to comparable counts in matched control cows. Mild cases of mastitis were preceded by higher cell counts than were found in control cows but the same phenomenon was not observed in acute cases of mastitis. Both mild and acute cases were followed by higher cell counts than were found in control cows.     

Detection of Replicative Form of HCV RNA in Peripheral Blood Leukocytes and Its Clinical Significance

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Comparison of selenium levels and glutathione peroxidase activity in bovine whole blood

Blood glutathione peroxidase activity and selenium levels were found to correlate well, indicating that glutathione peroxidase activity can be used to assess blood selenium levels in beef cattle. The glutathione peroxidase activity of blood is less stable than is the selenium concentration but when blood was stored at 4 degrees C, the glutathione peroxidase activity remained constant for seven days.