猪RBP1和RBP4基因的定位、组织表达谱分析、多态研究及相关分析

类维生素A（维生素A及其代谢产物）在动物的视觉、生长、繁殖、细胞的分化及胚胎发育等方面起重要的作用。利用猪的辐射杂种克隆板，将猪细胞视黄醇结合蛋白基因1（RBP1）和猪血浆视黄醇结合蛋白基因4（RBP4）分别定位在猪13号和14号染色体上。利用半定量RT－PCR方法，对这两个基因在成年五指山猪十二种不同组织（肺、骨骼肌、脾、心脏、胃、大肠、淋巴结、小肠、肝、大脑、肾、脂肪）的组织表达谱进行了分析。在猪的血浆视黄醇结合蛋白基因4中，发现一个单核苷酸多态位点（RBP4-A/G156），并用基于聚合酶链式反应的限制性片段长度多态性方法(PCR-RFLP)，对这一多态在莱芜黑猪、五指山小型猪、贵州小型猪、广西巴马小型猪和通城实验猪群中的分布进行了研究。在通城实验猪群中，进行了不同基因型与性状间的关联分析，发现其不同的基因型与胴体直长、胴体斜长、血红蛋白浓度、平均血细胞血红蛋白量、达上市体重日龄高度相关。在猪的生产和育种中，这一多态位点可能会成为有用的分子标记。     

母乳中VA与婴幼儿健康研究进展

母乳中含有多种VA,如视黄醇、β-胡萝卜素、β-隐黄质、α-胡萝卜素、番茄红素及叶黄素等,且母乳初乳、过渡乳和成熟乳中VA的含量不同,但总体上VA含量随着泌乳期的延长而降低。如母乳中含量最高的VA视黄醇,其在初乳、过渡乳、成熟乳中的含量分别为478～1 920、440～1 270、178～825 μg/L。母乳中VA含量除受泌乳时间影响外,还与地域、乳母膳食及母乳脂肪含量等有关。本文重点介绍不同国家/地区母乳中VA的种类、含量、影响因素及其健康作用（视觉健康、免疫健康和生长发育等）,以期为婴幼儿配方乳粉的发展提供参考。     

A comparison of retinol, retinal and retinyl ester concentrations in larvae of Atlantic halibut (Hippoglossus hippoglossus L.) fed Artemia or zooplankton

Atlantic halibut (Hippoglossus hippoglossus L.) larvae were fed enriched Artemia or zooplankton in duplicate tanks from 0 to 60 days after first‐feeding. Both diets and the larvae were analysed for vitamin A (VA) in order to confirm earlier findings, in which Artemia fed larvae had lower levels of VA compared with larvae fed zooplankton. Furthermore, we wanted to investigate the composition of the retinoids in the larvae. The results showed that Artemia and zooplankton contains low levels of VA, probably too low to sustain the assumed requirement. Nevertheless, larvae fed Artemia had the same level of retinal and retinol as larvae fed zooplankton. We found a significant lower level of retinyl esters in larvae fed Artemia. The total VA level was lower in larvae fed Artemia only at the end of the feeding trial after the onset of metamorphosis. Our conclusion is that feeding Artemia to Atlantic halibut larvae is not likely to cause VA deficiency.     

Association of RBP‐4 gene polymorphisms with follicular cysts in large white sows

Follicular cysts, which is a common infertility disease, can cause financial losses in pig breeding programmes. The pathogenesis and mechanisms of the formation of follicular cysts are not understood clearly. In our previous study, the concentration of retinol‐binding protein 4 (RBP‐4) in the follicular fluid (FF) of the ovary with follicular cysts was found to be significantly higher than that of normal ovary, thereby suggesting that RBP‐4 may be a candidate biomarker for porcine follicular cysts. To study the association of RBP‐4 and follicular cysts further, we detected the polymorphisms of the RBP‐4 gene and the presence of follicular cysts by PCR‐Restriction fragment length polymorphism (RFLP) assay. In this study, we screened the mutations of RBP‐4 gene in 79 sows with follicular cysts and 100 normal sows without cysts. Results showed that +249‐63G>C polymorphisms were significantly associated with follicular cysts, and sows with CC genotype in RBP‐4 gene had a high risk of developing follicular cysts. Hence, our findings further proved that RBP‐4 may be a novel biomarker for follicular cysts, which may be valuable for the diagnosis of follicular cysts and molecular breeding of pigs.     

Apelin-13对小鼠血脂代谢的影响及其机制研究

本研究旨在探索Apelin-13调控小鼠血脂代谢的生理机制。试验选取体重相近的小鼠96只,随机分为4个处理,每个处理4个重复。对照组和3个试验组小鼠分别每天皮下注射100 μL 0.9%的生理盐水、10、20和50 μg·kg^-1的Apelin-13,试验期为14 d。测定小鼠的生长性能、血清Apelin-13和血脂含量,并采用Illumina Novaseq测序平台进行小鼠肝组织转录组测序（RNA-seq）。结果表明,与对照组相比,皮下注射Apelin-13极显著提高了血清Apelin-13和高密度蛋白胆固醇含量（P<0.01）,显著降低了血清甘油三酯含量（P<0.05）。试验组和处理组共检测到340个上调和350个下调的差异表达基因（differentially expressed genes,DEGs）。这些DEGs最显著富集的信号通路为视黄醇代谢,该通路中的细胞色素P450（Cyp）和尿苷二磷酸葡萄糖醛酸转移酶（Ugt）两个超家族基因大部分上调表达。试验结果表明,Apelin-13可能通过调控视黄醇代谢信号通路中的Cyp和Ugt两个超家族基因的表达来调控小鼠的血脂代谢。     

Blood biochemical values in Japanese Black breeding cows in Kagoshima Prefecture,Japan

To obtain blood biochemical basic data of Japanese Black breeding cows in Kagoshima Prefecture, Japan, blood samples were obtained from 857 clinically healthy multiparous cows on 71 farms. Cows were divided into three stages: the prepartum stage (between 20 and 80 days before parturition, n=290), lactation stage (between 20 and 80 days after parturition, n=283) and maintenance stage (between 120 and 200 days after parturition, n=284). The mean concentration of total protein and total cholesterol, and the mean activities of aspartate aminotransferase in the lactation stage tended to be higher than those in the prepartum and maintenance stages. The mean concentration of glucose in the prepartum stage tended to be lower than that in the lactation and the maintenance stages. The mean concentration of nonesterified fatty acids and beta-hydroxybutyrate in the prepartum stage tended to be higher than those in the lactation and maintenance stages. The mean concentration of serum retinol was approximately 30 µg/dl in all stages. These results suggest that the blood biochemical values of multiparous Japanese Black cows vary with breeding stages, and it is considered that the blood parameters obtained in this study are useful as indices for health management of Japanese Black breeding cows.     

Concentrations of retinol and tocopherols in the milk of cows supplemented with conjugated linoleic acid

This study was performed to investigate the hypothesis that supplementation of conjugated linoleic acid (CLA) changes the concentrations of retinol and tocopherols in the milk of cows. To investigate this hypothesis, Holstein cows received daily from 3 weeks ante‐partum to 14 weeks post‐partum either 172 g of a CLA‐free rumen‐protected control fat (control group, n = 20) or the same amount of a rumen‐protected CLA fat, supplying 4.3 g of cis‐9, trans‐11 CLA and 3.8 g of trans‐10, cis‐12 CLA per d (CLA group, n = 20). Milk samples (collected at weeks 1, 3, 5, 8 and 11 of lactation) were analysed for retinol, α‐ and γ‐tocopherol concentrations. Milk of cows supplemented with CLA had higher concentrations of retinol (+34%), α‐tocopherol (+44%) and γ‐tocopherol (+21%) than milk of control cows (p < 0.05). The daily output of these vitamins via milk was also greater in cows of the CLA group than in cows of the control group (+36, 50 and 24% for retinol, α‐tocopherol and γ‐tocopherol, respectively, p < 0.05). In agreement with higher concentrations of tocopherols, concentrations of thiobarbituric acid‐reactive substances, determined in milk of week 5, were lower in cows of the CLA group than in control cows, indicative of a lower susceptibility of milk lipids to peroxidation. Plasma concentrations of retinol and α‐tocopherol, determined at 1 and 5 weeks post‐partum, were not different between the two groups of cows. In conclusion, this study shows that supplementing dairy cows with a moderate amount of CLA causes an increase of the concentrations of vitamins A and E in the milk and results in an increased output of those vitamins via milk. These effects might be beneficial with respect to the nutritional value of dairy products and the susceptibility of milk fat to oxidative deterioration.     

Estimation of dietary vitamin A requirement of juvenile soft‐shelled turtle,Pelodiscus sinensis

A feeding trial was conducted to evaluate the dietary vitamin A requirement of the soft‐shelled turtle Pelodiscus sinensis. Turtles with an approximate body weight of 6.8 g were fed diets containing 0–4.58 mg kg^?1 (15 251 IU) vitamin A (retinyl acetate) for 8 weeks. Turtles fed diets without vitamin A showed the lowest growth performance. The animals gradually gained weight when dietary vitamin A increased from 0 to 3.6 mg kg^?1. Liver vitamin A gradually increased with supplementation of dietary vitamin A of up to 2.7 mg kg^?1. Beyond this concentration, the vitamin A in the liver significantly increased. Data analysis using a polynomial regression or exponential model showed that the estimated dietary vitamin A requirement for juvenile soft‐shelled turtles based on weight gain, red blood cell count and liver vitamin A measurements was 3.48, 2.84 and 2.58 mg kg^?1, respectively.     

视黄醇对仔猪睾丸支持细胞体外生物学特性的影响

支持细胞是构成曲细精管上皮的重要组成细胞,视黄醇及其衍生物是雄性动物睾丸发育和精子发生中所必须的物质,支持细胞是睾丸内视黄醇及其衍生物作用的靶细胞。本研究以3~5周龄仔猪睾丸支持细胞为材料,通过分析视黄醇对体外支持细胞活力、增殖和凋亡相关基因表达、相关细胞因子转录及合成的影响,评价视黄醇对支持细胞体外生物学特性的影响。结果显示,与对照组相比,视黄醇添加浓度达到5.000、0.125 μmol·L^-1时,显著抑制培养24 h和72 h的细胞活性(P<0.05);添加1.250 μmol·L^-1视黄醇组和对照组相比,增殖相关基因PCNA、BCL-2和C-MYC显著下调(P<0.05),凋亡相关基因BAX显著上调(P<0.05),且支持细胞内GDNF、CSF-1和EGF在基因表达和蛋白质水平上均显著降低(P<0.05)。体外培养条件下,添加视黄醇抑制支持细胞活性,促进细胞凋亡。     

Hyperostotic polyarthropathy in a rabbit - a suspected case of chronic hypervitaminosis A from a diet of carrots

Chronic hypervitaminosis A can occur in many species after excessive dietary intake of Vitamin A (retinol). The most common presentation of chronic hypervitaminosis A is a polyarthropathy with hyperostosis and ankylosis of various joints. This case report describes a probable case of naturally occurring hypervitaminosis A-induced polyarthropathy in a rabbit after chronic ingestion of a diet made up almost exclusively of carrots. Carrots do not contain retinol, but are rich in provitamin A (or beta-carotene). Rabbits are unique in that they can convert 100% of dietary beta-carotene into retinol. A syndrome of naturally occurring hypervitaminosis A-induced polyarthropathy has not been described in a rabbit before.