牛角地黄汤对α-溶血素诱导红细胞溶血的抑制作用研究

[目的]牛角地黄汤是中国传统医学中用于清热解毒凉血化瘀的基础方剂,由水牛角、地黄、赤芍、丹皮、钩藤等主要成分组成。本研究旨在研究分析牛角地黄汤及方剂中主要成分对金黄色葡萄球菌α-溶血素诱导兔红细胞溶血作用的影响。[方法]将牛角地黄汤水煎剂及其各单味中药的水煎剂分别与金黄色葡萄球菌溶血毒素作用不同时间后检测溶血活性。[结果]研究发现,无论与溶血素同时给药或是在溶血素染毒之前给药,亦或是与溶血素预先单独作用一段时间后再给药,牛角地黄汤煎剂均能抑制α-溶血素致兔红细胞破裂的溶血作用。同时还发现,方剂中的赤芍和丹皮水煎液对溶血素破坏兔红细胞致溶血的活性抑制作用最为明显,是牛角地黄汤中起主要抗溶血作用的中药组分。而牛角和地黄的水煎液在各试验条件下均未表现出抑制溶血素溶血的作用。[结论]本研究表明,牛角地黄汤具有显著的对抗溶血素诱导红细胞溶血的作用,方中起主要作用的成分是赤芍和丹皮,钩藤的这种作用则比较弱,而牛角和地黄则不是牛角地黄汤中抗溶血作用的主要成分。     

Effect of MK‐467 on organ blood flow parameters detected by contrast‐enhanced ultrasound in dogs treated with dexmedetomidine

ObjectiveTo evaluate the dexmedetomidine‐induced reduction in organ blood flow with quantitative contrast‐enhanced ultrasound (CEUS) method and to observe the influence of MK‐467 on such reduction.Study designRandomized cross‐over study.AnimalsSix adult purpose‐bred laboratory beagle dogs (mean body weight 15.3 ± 1.9 kg).MethodsContrast‐enhanced ultrasound was performed on six conscious healthy laboratory beagles. The animals on separate occasions underwent three treatments: awake without any medication (CTRL), dexmedetomidine 10 μg kg^?1 (DEX) and DEX + MK‐467 500 μg kg^?1 (DMK) intravenously (IV). The kidney (10–15 minutes post‐treatment), spleen (25–30 minutes post‐treatment), small intestine (40–45 minutes post‐treatment) and liver (50–55 minutes post‐treatment) were examined with CEUS. A time curve was generated and the following perfusion parameters were analysed: arrival time (AT), time to peak from injection (TTPinj), peak intensity (PI) and wash‐in rate (Wi). In addition to CEUS, renal glomerular filtration rate was indirectly estimated by the rate of iohexol elimination.ResultsAT and TTPinj were significantly higher for DEX than for CTRL in all studied organs. The same parameters were significantly higher for DEX than for DMK in the kidney, spleen and small intestine. PI was significantly lower for DEX than for CTRL or DMK in the kidney. Wi was significantly lower for DEX than for CTRL or DMK in the kidney and significantly lower than for CTRL only in the small intestine. Plasma concentration of iohexol was significantly higher after DEX than CTRL administration.ConclusionsContrast‐enhanced ultrasound was effective in detecting DEX‐induced changes in blood flow. MK‐467 attenuated these changes.Clinical relevanceClinicians should consider the effects of the sedation protocol when performing CEUS. Addition of MK‐467 might beneficially impact the haemodynamic function of sedation with alpha‐2 adrenoceptor agonists.     

家蚕整合素基因Bmintegrin αPS3的鉴定及亚细胞定位

【目的】克隆家蚕整合素基因Bmintegrin αPS3,分析其mRNA表达及亚细胞定位特征,并进行原核表达及蛋白纯化,为进一步探究Bmintegrin αPS3在家蚕中的功能奠定基础。【方法】利用RACE方法克隆获得Bmintegrin αPS3的cDNA全长序列;采用RT-PCR方法检测Bmintegrin αPS3的时空表达;在原核表达系统中表达获得Bmintegrin αPS3重组蛋白;并构建Bmintegrin αPS3真核表达载体,转染家蚕胚胎细胞系分析其蛋白亚细胞定位。【结果】家蚕整合素基因Bmintegrin αPS3 编码框全长为2 895 bp,编码965个氨基酸,含有整合素家族蛋白典型的integrin alpha结构域,预测其为跨膜蛋白。构建了Bmintegrin αPS3的原核表达系统,并利用亲和层析纯化获得高纯度的Bmintegrin αPS3原核表达蛋白,为进一步制备抗体获得足够的抗原。在家蚕细胞系中外源表达Bmintegrin αPS3蛋白,显示其主要定位于细胞质和细胞膜中。另外,RT-PCR结果显示Bmintegrin αPS3 mRNA 在血细胞中为特异持续表达。【结论】获得了Bmintegrin αPS3的完整cDNA序列及表达特征,经原核表达、纯化获得其融合蛋白,在细胞水平上初步分析了其亚细胞定位情况。     

ORIGINAL ARTICLE: 2‐Oxoglutaric acid administration diminishes fundectomy‐induced osteopenia in pigs

Partial surgical removal of the stomach (fundectomy, FX) leads to osteopenia in animals and humans. FX adversely affects the bone. 2‐oxoglutaric acid is a precursor of glutamine and hydroxyproline – the most abundant amino acid of collagen. The aim of the study was to investigate the effects of 2‐oxoglutaric acid on FX‐evoked osteopenia in pigs. Eighteen castrated male pigs of the Pu?awska breed were used. Twelve pigs were subjected to FX and divided into two groups: FX + AKG (the AKG group; AKG at the daily dosage of 0.4 g/kg of body weight) and FX + Placebo (the FXC group; received CaCO₃ as placebo). Remaining six pigs were sham‐operated (the SHO group). The pigs were euthanized at the age of 8 months and long bones were collected. Area bone mineral density (aBMD) and bone mineral content (BMC) were measured; morphology, geometry and biomechanical properties were determined. Moreover, the serum concentrations of selected hormones and one marker of bone metabolism were determined. FX caused osteopenia in the pigs and treatment with AKG greatly reduced these effects of FX in pigs. Negative effect of fundectomy on the skeletal system leading to decreased bone mass in pigs is associated with lowered body gain and activity of the gastric–hypothalamic–pituitary axis. Better definitions of each of the local and systemic hormonal and structural components associated with fundectomy‐induced decreased bone mass that separately and together determine the whole bone properties may lead to identify opportunities for prevention.     

28种牧草的脂肪酸组成分析研究

对多花黑麦草等28种牧草样品进行了脂肪酸组成的测定,并进行分析比较。结果表明,α-亚麻酸是牧草中含量最高的脂肪酸,占脂肪酸组成的43.78%。多不饱和脂肪酸占牧草脂肪酸组成的54.71%,占不饱和性脂肪酸的95.18%。说明牧草的不饱和性脂肪酸主要是二键以上的多不饱和脂肪酸。禾本科牧草中,多花黑麦草及狼尾草属的牧草α-亚麻酸含量较高,其中多花黑麦草69.9%;象草(N51)67.6%;红象草62.2%;杂交狼尾草61.0%。禾本科与豆科牧草的α-亚麻酸含量差异不显著。     

The effect of tree cover on the biomass and diversity of the herbaceous layer in a Mediterranean dehesa

Mediterranean dehesas are an archetypal example of high nature value farmland in Europe. To better understand the interactions between trees and grasslands in the conservation and management of the dehesa, we measured, over 3 years, the contribution of scattered trees to the diversity and biomass dry‐matter (DM) yield of the herb layer and the relationship between the herb layer (biomass and diversity interaction) and the trees. The study was based on 135 subplots representing four aspects and three distances from fifteen trees selected randomly in a plot within a dehesa in central Spain. The specific questions were the following: (i) To what extent do trees affect biomass of the herbaceous layer? (ii) What is the relationship between herbaceous layer biomass and species diversity under the influence of trees? and (iii) What are the effects of trees and small‐scale topography on the herbaceous diversity? Results differed between years depending on water availability. Legume and forb DM yields were highest at the edge of the tree crown and beyond the canopy. Species richness was positively related to legume DM yield, and both were higher at the bottom of the hillside, especially in dry conditions. Given the importance of tree influence on the biomass and diversity of grasslands, maintaining proper agroforestry management practices for trees and grasslands is important for conserving dehesa diversity and productivity under the conditions of high climatic and soil variability in Mediterranean dehesas.     

Effects of alpha-lipoic acid supplementation on antioxidative ability and performance of sows and nursing piglets

The current study was carried out to determine the effects of alpha‐lipoic acid (LA) supplementation during late‐gestation and lactation on antioxidative ability and performance of sows and their nursing piglets. A total of 160 multiparous sows were randomly allocated to four treatments with 40 replicates per treatment according to parity number and backfat (BF) thickness. Sows were fed 1 of 4 diets from day 85 of gestation to day 21 of lactation. Diets were control without LA; 400 ppm LA supplementation; 600 ppm LA supplementation; and 800 ppm LA supplementation. BF thickness of sows was determined on day 85 and 110 of gestation and days 1 and 21 of lactation. Piglet bodyweight was measured at birth, days 7, 14 and 21. Blood samples were obtained from the sows, and average daily feed intake (ADFI) of the sows during lactation was recorded. There were no differences in BF thickness or ADFI among treatment groups. Dietary LA supplementation resulted in a decrease in blood urea nitrogen (p < 0.01) concentration at days 110 of gestation. Dietary 800 ppm LA increased serum glutathione peroxidase (GSH‐Px) activity (p < 0.05) and reduced maleic dialdehyde levels (p < 0.01) of sows compared with the control diet at days 21 of lactation. Alpha‐lipoic acid supplementation increased the birthweight and weaning weight of piglets (p < 0.01) compared with the control group. Weight gains of piglets from sows fed the 800 ppm LA diets were greater (p < 0.01) between days 7 and 14 compared with piglets from control sows. Weight gains of piglets from sows fed the LA‐supplemented diets were greater between days 14 and 21 (p < 0.05) and between days 1 and 21 (p < 0.01) compared with piglets from control‐fed sows. In conclusion, the results indicate that antioxidant LA was effective in enhancing antioxidant enzymes activity and improving the performance of sows and their nursing piglets.     

Effect of yohimbine on detomidine induced changes in behavior,cardiac and blood parameters in the horse

ObjectiveTo describe selected pharmacodynamic effects of detomidine and yohimbine when administered alone and in sequence.Study designRandomized crossover design.AnimalsNine healthy adult horses aged 9 ± 4 years and weighing 561 ± 56 kg.MethodsThree dose regimens were employed in the current study. 1) 0.03 mg kg^?1 detomidine IV, 2) 0.2 mg kg^?1 yohimbine IV and 3) 0.03 mg kg^?1 detomidine IV followed 15 minutes later by 0.2 mg kg^?1 yohimbine IV. Each horse received all three treatments with a minimum of 1 week between treatments. Blood samples were obtained and plasma analyzed for detomidine and yohimbine concentrations by liquid chromatography-mass spectrometry. Behavioral effects, heart rate and rhythm, glucose, packed cell volume and plasma proteins were monitored.ResultsYohimbine rapidly reversed the sedative effects of detomidine in the horse. Additionally, yohimbine effectively returned heart rate and the percent of atrio-ventricular conduction disturbances to pre-detomidine values when administered 15 minutes post-detomidine administration. Plasma glucose was significantly increased following detomidine administration. The detomidine induced hyperglycemia was effectively reduced by yohimbine administration. Effects on packed cell volume and plasma proteins were variable.Conclusions and clinical relevanceIntravenous administration of yohimbine effectively reversed detomidine induced sedation, bradycardia, atrio-ventricular heart block and hyperglycemia.     

Regulation of COX-2 expression in canine prostate carcinoma: increased COX-2 expression is not related to inflammation

BACKGROUND: Cyclooxygenase-2 (COX-2) expression has been documented in human and canine prostate carcinoma (PCA). Canine PCA is a histologically heterogeneous tumor, sometimes including inflammatory infiltrates. However, it is unknown whether COX-2 expression in canine PCA is related to the histologic type of tumor, to the presence of inflammation, or to both. Moreover, little is known about the mechanisms regulating COX-2 expression in neoplastic tissue. HYPOTHESIS: COX-2 expression is related to the presence of inflammation in canine PCA and correlates with the degree of tumor differentiation. METHODS: The expression of COX-2 was examined in 28 cases of canine PCA by immunohistochemistry. In addition, a neoplastic and a nonneoplastic canine prostatic cell line were used to investigate the effects of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), phorbol 12-myristate 13-acetate (PMA), epithelial growth factor (EGF), and specific signal transduction pathway inhibitors on COX-2 expression. RESULTS: Twenty-four of the 28 prostate tumors showed COX-2 expression. The presence of inflammatory infiltrates in tumor tissue was associated with lower COX-2 expression scores. In vitro, TNF-alpha, IL-6, and EGF increased COX-2 expression in nonneoplastic cells but not in PCA cells, where baseline expression was high. COX-2 expression in PCA cells could be suppressed by means of specific phosphatidyl inositol-3 kinase (PI3K), protein kinase C (PKC), or inhibitor of extracellular signal-related kinase (ERK/MAPK) inhibitors. CONCLUSIONS AND CLINICAL IMPORTANCE: COX-2 is expressed in canine PCA; however, expression is not related to the presence of inflammatory infiltrates. This conclusion is further supported by the finding that the cytokines TNF-alpha and IL-6 and their involved signaling pathways do not stimulate COX-2 expression in malignant canine prostate cells.     

重组胸腺素α1在大肠杆菌中的表达

重组胸腺素alpha 1(Tα1)基因在大肠杆菌中进行表达。根据大肠杆菌密码子偏爱性合成Tαl基因并连接到pMD18-T载体上,双酶切回收的Tαl插入到pET32a后,转化大肠杆菌BL21,IPTG诱导,经15%SDS-PAGE检测,胸腺素αl获得可溶性表达。检测到与目的蛋白相对分子量(21.8KDa)相符的条带。重组胸腺素alpha 1在大肠杆菌中获得可溶性表达。