Detection of γ-H2AX,a Biomarker for DNA Double-strand Breaks,in Urinary Bladders of N -Butyl- N -(4-Hydroxybutyl)-Nitrosamine-Treated Rats

To evaluate the potential role of DNA repair in bladder carcinogenesis, we performed an immunohistochemical analysis of expression of various DNA repair enzymes and γ-H2AX, a high-sensitivity marker of DNA double-strand breaks, in the urothelium of male F344 rats treated with N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN), a bladder-specific carcinogen. Our results clearly demonstrated that γ-H2AX aggregation was specifically generated in nuclei of bladder epithelial cells of BBN-treated rats, which was not found in untreated controls or mesenchymal cells. γ-H2AX-positive cells were detected not only in hyperplastic and neoplastic areas but also in the normal-like urothelium after BBN treatment. These data indicate that γ-H2AX has potential as a useful biomarker for early detection of genotoxicity in the rat urinary bladder. To the best of our knowledge, this is the first report demonstrating expression of γ-H2AX during bladder carcinogenesis.     

Effect of intake of linoleic acid and α‐linolenic acid levels on conversion into long‐chain polyunsaturated fatty acids in backfat and in intramuscular fat of growing pigs

A study was conducted to determine the effect of two levels of linoleic acid (LA) intake at either high or low α‐linolenic acid (ALA) intake on their conversion and subsequent deposition into long‐chain (20–22 C‐atoms) polyunsaturated fatty acids (LC PUFA) in muscle and backfat in growing pigs. In a 2 × 2 factorial arrangement, 32 gilts from 8 litters were assigned to one of four dietary treatments, varying in LA and ALA intakes. Low ALA and LA intakes were 0.15 and 1.31 g/(kg BW^0.75/day), respectively, and high ALA and LA intakes were 1.48 and 2.65 g/(kg BW^0.75/day) respectively. There was a close positive relation between intake of ALA and the concentration of ALA in backfat and in intramuscular fat. Dietary ALA did not affect the concentration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), but increased docosapentaenoic acid (DPA) in backfat. High ALA intake did not significantly affect DHA but significantly increased EPA, 20:3 n‐3 and DPA concentrations in intramuscular fat. The n‐3 LC PUFA proportion in backfat was increased from approximately 1–3%, which may be useful to enrich meat with these fatty acids. The effect of ALA intake on n‐3 LC PUFA was suppressed by LA intake. Dietary ALA suppressed the concentration of n‐6 LC PUFA in blood plasma by more than 50%. When compared at equal incremental dose, the inhibiting effect of ALA on blood arachidonic acid was stronger than the stimulating effect of LA as precursor.     

Trade-off expression of melatonin receptor subtypes (Mel1a and Mel1b) and androgen receptor in lung of a tropical bird,Perdicula asiatica

The role of circulatory steroid hormone along with melatonin in lung of any seasonally breeding bird has never been explored so far. This could be interesting because steroid hormones are immunosuppressive while melatonin is immunostimulatory in nature. In our present study, we report the effect of exogenous melatonin and testosterone on expression of melatonin receptor subtypes (Mel_1a and Mel_1b) and androgen receptor in lung of a tropical bird Perdicula asiatica. Birds were collected from vicinity of Varanasi and acclimatized in laboratory with sufficient food and water. The birds were treated with melatonin and testosterone at dose of 25 µg/100 g B.wt./day and 1 mg/100 g B.wt./day, respectively, for 28 days. At the end of the experiment, the birds were sacrificed and lung tissue and blood sample were collected for immunohistochemistry, Western blot analysis and hormonal assay. Testosterone treatment increased circulatory testosterone and upregulated expression of androgen receptors whereas downregulated expression of melatonin receptor subtypes Mel_1a and Mel_1b. Melatonin administration increased peripheral melatonin and upregulated expression of melatonin receptor subtypes Mel_1a and Mel_1b while downregulated androgen receptor. Thus, our results suggest that a trade-off relationship between melatonin and testosterone exists in regulation of their receptors in lung of Perdicula asiatica.     

不同CO2浓度对苜蓿分枝期光合性能的影响

以苜蓿（Medicago sativa）‘Maverick’为试验材料,研究不同CO₂浓度下苜蓿分枝期的光合能力日变化情况,探索未来大气CO₂浓度上升过程中苜蓿的生产潜力,为未来牧草种植产业提供指导。结果表明：当开顶式生长室中CO₂浓度为550和700 μmol·mol^-1时,‘Maverick’的叶绿素含量对比室外对照组分别提高了2.4%和3.6%,但差异不显著。净光合速率（Pn）日平均值分别提高了114.69%和116.54%。此外,CO₂浓度升高提高了苜蓿的光系统Ⅱ（PSII）的原初光能转换效率（Fv/Fm）、潜在活性（Fv/Fo）、实际光化学量子产量（Yield）,并显著增加了光化学淬灭（qP）,降低非光化学淬灭（NPQ）。研究结果表明在高CO₂浓度条件下更利于‘Maverick’叶绿素捕获光能,PSII反应中心比例大,光合作用增强。     

马头山羊线粒体细胞色素b基因遗传多样性研究

测定了马头山羊品种16个个体的细胞色素b基因全序列(1 140 bp),比较分析了群体中细胞色素b基因的碱基组成和序列间碱基的变异情况.结果显示:在该品种(群体)中细胞色素b基因序列中8个变异位点上观察到23次T-C间碱基转换,有11次T-G间碱基颠换发生在密码子第2位点,为非同义突变;观察到6种单倍型,单倍型多样度为0.808,核苷酸多样度为0.002 43.以绵羊为外群构建系统发生树(NJ tree).结果显示:马头山羊有两个母系起源,其中支系A占75%(12/16),支系B占25%(4/16).     

香猪睾丸miR-449b和miR-222分子的发育性变化

为了阐明miR-449b和miR-222对贵州从江香猪睾丸发育的调节作用。运用qPCR方法检测香猪睾丸组织中miR-449b和miR-222表达量的发育性变化,通过miRSystem数据库、Kobas数据库对预测的靶基因进行信号转导通路富集分析。经克隆测序,得到香猪miR-222序列,与猪的相似性为100%,而且物种之间miR-449b和miR-222两个分子的保守性都很高。香猪睾丸中miR-449b于1~4月龄期间的表达量逐渐上升,4月龄时表达量达到峰值,之后下降;miR-222的表达量呈现相反的变化趋势,于1~4月龄期间表达量逐渐下降,1月龄时最高,4月龄时最低,之后的表达量有所回升;已知物种的miR-449b和miR-222序列高度保守;经预测miR-449b靶基因有420个,富集于卵母细胞减数分裂、Notch信号通路、Ras信号通路等100个途径(P0.05),miR-222的靶基因有429个,富集于细胞周期、Fox O信号通路、TGF-β信号通路等69个通路(P0.05);miR-449b和miR-222的靶基因共享的通路为Wnt信号通路、MAPK信号通路、雌激素信号通路等42个(P0.05)。     

Bactrian camel (Camelus bactrianus) integrins αvβ3 and αvβ6 as FMDV receptors: Molecular cloning, sequence analysis and comparison with other species

Integrins are heterodimeric adhesion receptors that participate in a variety of cell–cell and cell–extracellular matrix protein interactions. Many integrins recognize RGD sequences displayed on extracellular matrix proteins and the exposed loops of viral capsid proteins. Four members of the αv integrin family of cellular receptors, αvβ3, αvβ6, αvβ1 and αvβ8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro, and integrins are believed to be the receptors used to target epithelial cells in the infected animals. To analyse the roles of the αv integrins from a susceptible species as viral receptors, we have cloned Bactrian camel αv, β3 and β6 integrin cDNAs and compared them to those of other species. The coding sequences for Bactrian camel integrin αv, β3 and β6 were found to be 3165, 2289 and 2367 nucleotides in length, encoding 1054, 762 and 788 amino acids, respectively. The Bactrian camel αv, β3 and β6 subunits share many structural features with homologues of other species, including the ligand binding domain and cysteine-rich region. Phylogenetic trees and similarity analyses showed the close relationships of integrin genes from Bactrian camels, pigs and cattle, which are each susceptible to FMDV infection, that were distinct from the orders Rodentia, Primates, Perissodactyla, Carnivora, Galliformes and Xenopus. We postulate that host tropism of FMDV may in part be related to the divergence in integrin subunits among different species.     

Effect of dietary supplementation of fish oil for lactating sows and weaned piglets on piglet Th polarization

The present study was designed to investigate the effect of dietary fish oil supplementation on piglet T helper cells (Th) polarization in relation to its impact on piglet serum interferon γ (IFN-γ) and interleukin 10 (IL-10) concentrations and splenic expression of Th1/Th2 characteristic genes. The diets of 18 gestating sows were supplemented with 7% lard (C) (n = 10) or 7% fish oil (T) (n = 8) from 10 d before parturition to weaning. At weaning, a split plot experiment was designed, 56 piglets, 28 each from sows fed with fish oil diet or lard diet, were divided into four groups of 7 replicates (one female and one castrated male per replicate) based on both sow diet during lactation and post-weaning piglet diet (C had 7% lard and T had 7% fish oil): CC, CT, TC, TT, and were fed the 7% fish oil or lard diet from day 35 to day 70. Serum concentrations of IFN-γ and IL-10, and Th1/Th2 related genes expression levels in spleen were measured and analyzed. The results showed that piglets fed with fish oil diet during post-weaning tended to have higher serum IFN-γ/IL-10 ratio (P = 0.09) than lard diet fed piglets. Lactation fish oil feeding increased splenic IL-12b, IL-12 receptor β2 (IL-12Rβ2), IL-2 and IFN-γ genes expression (P < 0.05 or P < 0.01) and post-weaning fish oil feeding increased splenic IL-12b (P = 0.06), IL-2 (P < 0.01) and IFN-γ (P = 0.08) mRNA expression than that in lard diet fed piglets at the end of this experiment. On the other hand, IL-4 gene expression (P = 0.01) in spleen was lower in weaned piglet from fish oil diet fed sows than that from lard diet fed sows. However, post-weaning piglets fed fish oil diet had higher splenic IL-4 (P = 0.06), IL-6 (P < 0.01) and IL-10 (P = 0.05) mRNA abundances than that fed with lard diet. These results indicated that dietary fish oil during lactation could increase Th1 polarization and accelerate immune maturation; while 7% fish oil in weaned piglets' diet was likely to increase Th2 cytokines expression.     

鸵鸟红细胞免疫功能研究

应用红细胞Ｃ３ｂ受体花环（Ｃ３ｂＲＲ）和免疫复合物花环（ＩＣＲ）试验,证实鸵鸟红细胞表面存在补体Ｃ３ｂ受体（Ｃ３ｂｒｅｃｅｐｔｏｒ,Ｃ３ｂＲ）,表明红细胞免疫系统（Ｒｃｄｃｅｌｉｍｍｕｎｅｓｙｓｔｅｍ,ＲＣＩＳ）的概念亦适用于这种动物;鸵鸟红细胞不但有传统医学观念上的功能,而且还可借助ＣＲ１（ＣｏｍｐｌｅｍｅｎｔｒｅｃｃｐｔｏｒｔｙｐｅＩ）进行免疫粘附,并以此为基础,表现出各种重要的免疫功能。     

New alleles of chicken CD8 alpha and CD3d found in Chinese native and western breeds

Chinese native chicken breeds provide useful resources for the study of genetic diversity. In this study, the alleles of CD8 alpha and CD3d cDNA from Chinese native and introduced western breeds of chicken were analyzed at the sequence level. Six alleles were found, due to 13 amino acid replacements in the extracellular domain of the CD8 alpha sequence. There were four alleles detected in the Chinese strains, and alleles 5 and 6 were identified for the first time. Allele 6 was shared by Langshan, Beijing Fatty and Recessive White Feather chickens. Allele 2, found in the Bigbone strain, was the same as that found in the Leghorn strain H.B15.H7, and allele 3 in the Xianju breed was also the same as in the Leghorn strain H.B15.H12. Two Leghorn lines (RPL line 7 and AY519197) and the Camellia possessed an allele (alleles 1, 4 and 5), respectively, that was not found in the other lines. Nine out of 13 amino acid replacements were situated in the putative major histocompatibility complex (MHC) class I binding CDR1 (positions 30, 33 and 34), CDR2 (positions 58, 62, 63 and 65) and CDR3 (positions 90 and 106). Except for the Xianju breed, the CD8 alpha cDNA of Chinese native chicken breeds shared high homology. Two alleles were found in CD3d. Three additional nucleotides were found at positions 64, 65 and 66 in the newly discovered allele 2. This led to a difference of four amino acids (at residues 22, 23, 24 and 25) in the extracellular domain of CD3d cDNA from the Gushi, Recessive White Feather and ISA chickens compared with these of the White Leghorn and T11.15 (NM_205512). Five hybridoma clones (1C9, 1H5, 4B11, 6G5 and 13C5) against chicken CD8 alpha were generated by DNA immunization. Two monoclonal antibodies (mAbs), 6G5 and 4B11, showed reactivity to the splenocytes from five Chinese native chicken breeds, the Recessive White Feather chicken and the Leghorn (AY519197), while mAbs 1C9, 1H5 and 13C5 showed no reaction with these breeds.