Spatiotemporal heterogeneity of PPARγ expression in porcine uteroplacenta for regulating of placental angiogenesis through VEGF-mediated signalling

Non-infectious prenatal mortality severely affects the porcine industry, with pathological placentation as a likely key reason. Previous studies have demonstrated that peroxisome proliferator-activated receptor gamma (PPARγ) deficiency causes defects in the uteroplacental vasculature and induces embryonic losses in mice. However, its role in porcine placental angiogenesis remains unclear. In the present study, PPARγ expression was investigated in porcine uteroplacental tissues at gestational day (GD) 25, GD40 and GD70 via quantitative polymerase chain reaction (qPCR), Western blot and immunohistochemistry (IHC). Moreover, the roles of PPARγ in porcine placental angiogenesis were investigated using a cell model of porcine umbilical vein endothelial cells (PUVECs) to conduct proliferation, migration and tube formation assays in vitro and a mouse xenograft model to assess capillary formation in vivo. The results showed that PPARγ was mainly located in the glandular epithelium, trophoblast, amniotic chorion epithelium and vascular endothelium, as indicated by the higher expression levels at GD25 and GD40 than at GD70 in endometrium and by higher expression levels at GD40 and GD70 than at GD25 in placenta. Moreover, PPARγ expression was significantly downregulated in placenta with dead foetus. In PUVECs, knocking out PPARγ significantly inhibited proliferation, migration and tube formation in vitro and inhibited capillary formation in mouse xenografts in vivo by blocking S-phase, promoting apoptosis and downregulating the angiogenic factors of VEGF and its receptors. Overall, the spatiotemporal heterogeneity of PPARγ expression in porcine uteroplacental tissue suggests its vital role in endometrial remodelling and placental angiogenesis, and PPARγ regulates placental angiogenesis through VEGF-mediated signalling.     

Pharmacokinetics and bioavailability of valnemulin in broiler chickens

Wang, R., Yuan, L.G., He, L.M., Zhu, L.X., Luo, X.Y., Zhang, C.Y., Yu, J.J., Fang, B.H., Liu, Y.H. Pharmacokinetics and bioavailability of valnemulin in broiler chickens. J. vet. Pharmacol. Therap. 34 , 247–251. The objective of this study was to investigate the pharmacokinetics and bioavailability of valnemulin in broiler chickens after intravenous (i.v.), intramuscular (i.m.) and oral administrations of 10 mg/kg body weight (bw). Plasma samples were analyzed by high‐performance liquid chromatography–tandem mass spectrometry (HPLC‐MS/MS). Pharmacokinetic characterization was performed by non‐compartmental analysis using WinNonlin program. After intravenous administration, distribution was wide with the volume of distribution based on terminal phase(V_z) of 4.27 ± 0.99 L /kg. Mean valnemulin t_1/2β(h), Cl_β(L /h /kg), V_ss (L /kg) and AUC_(0–∞)(μg·h /mL) values were 2.85, 0.99, 2.72 and 10.34, respectively. After intramuscular administration, valnemulin was rapidly absorbed with a C_max of 2.2 μg/mL achieved at 0.43 h (t_max), and the absolute bioavailability (F) was 88.81%; and for the oral route the same parameters were 0.66 ± 0.15 μg/mL, 1.54 ± 0.27 h and 74.42%. A multiple‐peak phenomenon was present after oral administration. The plasma profile of valnemulin exhibited a secondary peak during 2–6 h and a tertiary peak at 32 h. The favorable PK behavior, such as the wide distribution, slow elimination and acceptable bioavailability indicated that it is likely to be effective in chickens.     

Association of polymorphisms for nuclear receptor coactivator 1 gene with egg production traits in the maternal line of Shaobo hens

1. The objectives of the study were to find polymorphic sites and elucidate the association between SNPs in the nuclear receptor coactivator 1 (NCOA1) gene and reproductive traits. 2. SNPs were detected by PCR-SSCP and DNA sequencing. Four SNPs were detected, including T10155007A, T10125838C, G10118492A and G10109315T. Three polymorphisms were associated with total egg production at the age of 300 d and the G10109315T polymorphism was associated with age at first egg. 3. In conclusion, the NCOA1 gene can be used as a molecular marker for reproductive traits in hens.     

中国荷斯坦牛乳蛋白分子遗传多态性和产奶性状相关性的研究

从北京两个主要牛场共采得１８７头中国荷斯坦奶牛的血样,提取基因组ＤＮＡ,通过ＰＣＲ－ＲＦＬＰ方法对Ｋａｐｐａ酪蛋白、Ｂｅｔａ乳球蛋白（βｌｇ）和Ａｌｐｈａ乳白蛋白（α－ｌａ）进行了基因型的鉴定,并结合产奶性状进行统计分析,结果表明,牛群中上述３种乳蛋白基因的基因频率分别为Ｋ－ｃｎＡ７９％,Ｋ－ｃａＢ２１％,β－ｌｇＡ４３％,β－ｌｇＢ５７％,α－ｌｇＢ１００％,Ｋ－ＣＮ和β－ｌｇ基因位眯对产奶量没     

鹦鹉热衣原体荧光环介导等温扩增（LAMP）检测方法的建立

鹦鹉热衣原体(Chlamydia psittaci)是一种人兽共患病原体,能引起自然疫源性衣原体病,目前广泛分布于世界各地。本研究应用环介导等温扩增技术(LAMP),针对鹦鹉热衣原体23S RNA基因序列设计引物,并进行筛选以及敏感性和特异性试验,建立了鹦鹉热衣原体恒温荧光扩增方法。该方法在63℃恒温条件下1 h内即可显示结果,操作简单、快速,特异性强,灵敏度可达100拷贝/μL,且与流产衣原体、动物布鲁氏菌、牛结核分枝杆菌、沙门氏菌、大肠杆菌、金黄色葡萄球菌无交叉反应;用国产仪器可直接通过检测荧光信号判读结果,既增强了准确性,也避免了开盖污染产生假阳性;应用建立的LAMP方法对实验室保存的临床DNA样品进行检测,发现检测结果与QPCR相同。该方法的建立弥补了传统检测技术的不足,为实现鹦鹉热衣原体的现场快速诊断提供了技术支持。     

畜牧兽医执法办案应注意的问题之我见

结合畜牧兽医执法工作实践,总结了10项执法工作中经常遇到的问题,并有针对性的提出了处理建议,以便为国内畜牧兽医执法人员正确执法办案提供参考。     

Green tea polyphenols supplementation alters immunometabolism and oxidative stress in dairy cows with hyperketonemia

Peripartal cows often experience negative energy balance, and are therefore prone to suffering from metabolic diseases such as hyperketonemia, which causes financial losses in dairy farms. This study aimed to investigate the effect of green tea polyphenol (GTP) supplementation during the periparturient period on production performance, oxidative stress and immunometabolism in dairy cows with hyperketonemia. One hundred Holstein cows were assigned to GTP (0.2 g/kg DM; n = 50) or control (without GTP; n = 50) group based on body weight, previous milk yield, and parity on d 15 before expected parturition. Subsequently, 10 cows with hyperketonemia were selected from each group, according to blood β-hydroxybutyric acid (BHBA) concentration between 1.2 and 2.9 mmol/L from 2 to 3 d postpartum. All cows were fed a close-up diet and a lactation diet with or without GTP supply from 15 d prepartum until 30 d postpartum. Milk and blood samples were obtained from 20 cows selected with hyperketonemia on 10, 20, and 30 d postpartum. Compared with control cows, greater milk yield and lower somatic cell count were observed in GTP cows. The GTP group had lower concentrations of BHBA, free fatty acids, cholesterol, triglyceride, reactive oxygen species, malondialdehyde, and hydrogen peroxide, greater concentrations of glucose, lower activities of aspartate aminotransferase, alanine aminotransferase, and glutamyl transpeptidase, alongside greater activities of superoxide dismutase, glutathione peroxidase, and total antioxidant capacity. Additionally, GTP supplementation up-regulated concentrations of interleukin-6 and interleukin-10, but down-regulated concentrations of tumor necrosis factor-α, interleukin-1β, interleukin-2, interleukin-8, and interferon-γ in plasma. Greater concentrations of plasma immunoglobulin G were also detected in the GTP group. Overall, the data suggested that GTP supplementation from 15 d prepartum to 30 d postpartum improved the milk yield and health status in cows with hyperketonemia during early lactation.     

牛支原体和牛病毒性腹泻病毒二重二温式PCR检测方法的建立

为建立一种牛支原体(Mycoplasma bovis,MB)和牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)的快速鉴别诊断方法,针对MB的uvr C基因和BVDV的5'端非编码区(5'-UTR)保守基因序列,分别设计两对特异引物,并将三温式PCR扩增程序简化为二个温度梯度,建立了鉴别MB和BVDV的二重二温式PCR方法。该方法能同时扩增MB和BVDV,扩增产物大小分别为412和170 bp。特异性试验结果显示,该方法对参试的所有毒株只扩增MB和BVDV基因组,对其它牛病原体无扩增;敏感性试验结果显示,该方法最低能同时检测到104拷贝的两种目的核酸;干扰性试验结果显示,该方法能同时检测两个模板不同浓度的组合,试验结果不受模板影响。综上,本研究所建立的二重二温式PCR方法特异、敏感、快速、简便,可应用于MB和BVDV临床鉴别诊断和流行病学调查。     

Pharmacokinetics of sanguinarine,chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration

Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit®. However, little information is available on the pharmacokinetics of Sangrovit® in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit® and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half‐life (t_1/2) values of 1.05 ± 0.18 hr and 0.83 ± 0.10 hr for SA and DHSA, respectively. The maximum concentration (C_max) of DHSA (2.49 ± 1.4 μg/L) was higher that of SA (1.89 ± 0.8 μg/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 ± 5.4 and 6.08 ± 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 ± 0.63 (L·h^?1·kg^?1) with a t_1/2 of 0.34 ± 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 ± 1.05 and 0.52 ± 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit® had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit® after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.     

宿主域扩大的重组救活昆虫杆状病毒表达载体的构建及外源基因的表达

通过克隆的家蚕核多角体病毒解旋酶基因DNA与重组救活可线性化的苜蓿尺蠖核多角体病毒基因工程载体病毒 (BacPAK6 )DNA在昆虫细胞中发生重组 ,经累代筛选获得了既可感染家蚕又可感染秋粘虫细胞Sf 2 1的宿主域扩大的昆虫杆状病毒表达载体 (HyBacPAK6 )。HyBacPAK6DNA经Bsu36Ⅰ酶切后与含植酸酶基因的转移载体pVL1393 phy在家蚕细胞中重组后 ,通过蓝白斑筛选发现重组率可达 90 %以上。然而以杂交病毒为载体的外源基因表达量仅为以BmNPV为载体病毒的表达量的 2 0 %左右。分析认为HyBacPAK6可用于表达一些对蚕体有害的外源基因。