聚丙烯酰胺凝胶圆盘电泳分离奶牛血清碱性磷酸酶同功酶及鉴定组织来源

用聚丙烯酰胺凝胶圆盘电泳分离黑白花奶牛血清AKP同功酶酶谱及鉴定其组织来源。健康牛血清有5条酶带,以其泳动速率分为快(SF)、慢-1、慢-2、慢-3和慢-4(SS-1～4)。所有血清中除有主带SF和SS-1外,尚有1～3条慢带。骨只有一条泳动速率与血清主带SS-1相同的带,被认为是SS-1的来源。肝有2条泳动较快的带(LF-1和LF-2)和一条慢带LS。其中LF-1为窄的黄带,血清无对应带;LF-2与血清主带SF相对应,为SF的来源;LS为一条与SS-1相对应的浅而宽的带,为血液污染所致。小肠有3条窄的快带(IF-1～3)和一条宽的慢带(IS),慢带与比它稍快的骨带有部分重合出现于部分血清中,而3条快带未在血清出现。各种组织的AKP有不同的热稳定性。总之,血清主带SF和SS-1分别来源于肝和骨,SS-2来源于小肠,SS-3和SS-4在所分析的组织中无相应酶带。本研究较详细地描述了一种被改进的电泳方法,使其易于应用。     

Bone metabolic effects of single-dose zoledronate in healthy dogs

Zoledronate, an aminobisphosphonate with potent antiresorptive activity in bone, has the potential for treatment of malignant osteolysis and hypercalcemia in dogs. The purpose of this study was to evaluate the bone metabolic effects of a single dose of zoledronate in healthy dogs. Four skeletally mature, male, intact dogs received a 15-minute i.v. infusion of zoledronate at a dosage of 0.25 mg/kg. Urine N-telopeptide of type I collagen (NTx) excretion decreased significantly from baseline by 76, 63, 77, and 73% on days 7, 14, 21, and 28, respectively (P < .0125). Serum bone-specific alkaline phosphatase (bALP) decreased significantly from baseline by 36 and 42% on days 21 and 28, respectively (P < .0125). No changes were detected in indices of calcium homeostasis (ionized calcium, intact PTH, or urine calcium excretion). Single-dose i.v. zoledronate at 0.25 mg/kg appears to inhibit homeostatic osteolytic activity in healthy, skeletally mature dogs. Prospective studies should assess repeated-dose safety and activity in healthy and diseased animals of various age groups.     

Italian University System has to Match Educational Needs in Adherence to European Standards and Professional Scenario

The effect of a water-soluble fraction (WSF) of a non-pathogenic strain of Mycobacterium phlei was studied in bovine subclinical mastitis (SCM) by measuring the myeloperoxidase and acid phosphatase enzyme levels in the milk leukocytes. Forty-five cows were divided into three equal groups. Group I, consisting of 15 healthy cows, served as the control, whereas groups II and III each contained 15 cows with subclinical mastitis on the basis of a positive reaction in the California mastitis test (CMT). The cows in group II received 100 microg of WSF in 5 ml sterile phosphate-buffered saline, pH 7.4 (PBS) once only, while those in group III received 5 ml sterile PBS daily for 7 days, both treatments being given by the intramammary route. Observations were made up to 30 days after treatment (AT). The CMT of the healthy milk was negative (0), whereas it ranged between 1 and 2 points in SCM. The somatic cell count (SCC) increased significantly (p < 0.05) on day 3, then fell steeply from day 7 up to day 30 AT in the cows in group II. A steady decrease in the total bacterial count (TBC) was observed in the group treated with WSF but the bacterial counts remained high in the groups treated with PBS. The mean acid phosphatase level was enhanced by 119% on day 3 AT in group II but only by 18.7% in the cows in group III. The mean myeloperoxidase level was enhanced by 100% in the cows in group II but only by 18% in those in group III on day 3 AT. This significant reduction in the bacterial load in infected cows caused by intramammary infusion of WSF may be due to activation of the microbicidal activity of the neutrophils, but this requires confirmation.     

鱼腥蓝细菌PCC 7120中两个磷酸酶N端结构域活性分析

为了研究鱼腥蓝细菌(A nabaena sp.)PCC 7120中两个PP2C类蛋白磷酸酶PrpJ1和PrpJ2的磷酸酶活性,将编码两个蛋白N端至跨膜区的基因片段重组到质粒中,转化大肠杆菌进行原核表达,获得了可溶性的PrpJ1up和PrpJ2up蛋白.并且以pNPP为底物测定了所得蛋白的磷酸酶活性,双倒数作图法结果显示PrpJ1up蛋白的KK为0.30 mmol/L,Vmax为7.10 nmol/min;PrpJ2up蛋白的Km为0.24 mmol/L,Vmax为0.43 nmol/min.     

氨氮对凡纳滨对虾稚虾的急性毒性作用

实验研究了氨氮对体长3．2cm、体质量0．356g（n=10）与体长1．6cm、体质量0．036g（n=15）两种规格凡纳滨对虾（Litopenaeus vannamei）稚虾的急性毒性效应。血氨和碱性磷酸酶活性的测定结果显示,血氨随氨氮浓度的升高而升高,碱性磷酸酶（AKP）活性随氨氮的升高而下降。急性毒性实验结果表明,体长3．2cm稚虾的游离态氨24hLG。为2．068mg／L、48hLC5。为1．654mg／L、96hLG。为1．395mg／L,生存安全浓度sc为0．3969mg／L;体长1．6em稚虾的游离态氨24hLGo为1．577mg／L、48hLG0为1．042mg／L、96hL氏为1．012mg／L,生存安全浓度Sc为0．2065mg／L。     

镜鲤碱性磷酸酶和酸性磷酸酶QTL 定位分析

磷酸酶普遍存在于动物的各组织中,对磷酸单酯键具有水解活性,是机体生长代谢、保持内环境稳定和维持机体健康所必需的酶。以镜鲤(Cyprinus carpio L.)全同胞家系190个个体为材料,用992对微卫星(SSR)标记进行基因组扫描,采用区间作图法,对肠组织的碱性磷酸酶和酸性磷酸酶进行了QTL定位分析。QTL检测显示:5个QTL区间与酸性磷酸酶活性相关,其中3个QTL为99%染色体水平显著性,分别位于LG2(CA2049-CA2371)、LG25(HLJ2941-HLJ3946)和LG28(CA2263-HLJ2873),另外两个QTL区间为95%染色体水平显著性,位于LG14(HLJ3275-CA174)和LG14(CA1276-CA2208),解释表形变异率范围为8.1%～38.9%;2个与碱性磷酸酶相关的QTL区间均为95%染色体水平显著性,分别位于LG8和LG13,可解释变异率分别为7.3%和7.0%。     

两种方法所获鸭破骨细胞数量及活性比较

对直接从鸭骨髓腔机械分离的成熟破骨细胞（osteoclasts,OC）和由鸭骨髓来源单核细胞融合成的OC样多核巨细胞（multinucleatedgiantcells,MNGCs）进行培养,分别进行抗酒石酸酸性磷酸酶（tartrate-resistantacidphosphatase,TRAP）染色并计数,扫描电镜观察象牙片吸收陷窝,比较了2种方法获得0C的骨吸收功能。结果显示,2种方法均能分离培养出TRAP阳性且具有骨吸收功能的多核OC,但直接分离获得的成熟OC骨吸收功能更强。     

谷氨酰胺对断奶仔猪生长性能、营养物质表观消化率、空肠碱性磷酸酶活性及与肠道健康相关因子基因表达的影响

本研究的目的是探讨饲粮中添加谷氨酰胺对断奶仔猪生长性能、营养物质表观消化率、空肠碱性磷酸酶活性及与肠道健康相关因子基因表达的影响.将128头21日龄断奶的“杜×长×大”三元杂交仔猪按照体重、性别一致的原则随机分成2组,分别为对照组和添加1.00％谷氨酰胺组,每组4个重复,每个重复16头仔猪.在仔猪断奶后第10天和第30天,分别从每个重复中采集新鲜粪便样本,然后选择1头接近平均体重的仔猪进行屠宰,取空肠,刮取黏膜.试验期为30 d.结果表明:在断奶后第10天,与对照组相比,谷氨酰胺组平均日增重、干物质、粗蛋白质表观消化率和表观代谢能分别显著提高了19.05％、9.06％、4.77％和7.02％(P＜0.05),大多数必需氨基酸和非必需氨基酸表观消化率均有显著的提高(P＜0.05);在断奶后第30天,谷氨酰胺组干物质、粗蛋白质表观消化率和表观代谢能分别显著提高了5.90％、2.80％和6.43％ (P ＜0.05).在断奶后第10天和第30天,谷氨酰胺组仔猪空肠刷状缘碱性磷酸酶活性比对照组分别提高了30.09％(P＜0.05)和5.98％ (P ＞0.05).谷氨酰胺组空肠过氧化物酶体增殖体激活受体-γ和丙酮酸激酶基因mRNA表达水平在断奶后第10天比对照组分别下降了10.00％和30.00％ (P ＞0.05);在断奶后第30天比对照组分别显著下降了42.22％和44.52％ (P ＜0.05).在断奶后第10天和第30天,谷氨酰胺组空肠雷帕霉素靶蛋白基因mRNA表达水平分别比对照组提高了22.00％ (P ＞0.05)和22.38％ (P ＜0.05).以上结果提示,谷氨酰胺通过提高空肠碱性磷酸酶活性和调控与肠道健康相关因子基因表达水平来提高断奶仔猪的生长性能和营养物质表观消化率.     

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.