乙烯对匍匐翦股颖ISR抗病反应中AsA-GSH循环及胼胝质沉积的影响

以匍匐翦股颖品种“Penn-A4”为试验材料,侵染立枯丝核菌后,经丁二醇(BDO)诱导产生系统抗性(ISR),喷施不同浓度的乙烯合成抑制剂氯化钴(CoCl₂)和促进剂1-氨基环丙烷羧酸(ACC)后,检测抗坏血酸-谷胱甘肽(AsA-GSH)循环中抗坏血酸(AsA)和还原型谷胱甘肽(GSH)含量及相关酶活性的变化,并观察匍匐翦股颖ISR反应中胼胝质沉积的变化。结果表明,乙烯抑制剂处理下,匍匐翦股颖幼苗中AsA含量较低,抗坏血酸过氧化物酶(APX)活性下降,大量GSH被催化还原为氧化型谷胱甘肽(GSSG),GSSG大量积累,同时谷胱甘肽还原酶(GR)活性较低,GSSG经GR少量还原为GSH。乙烯促进剂处理下,AsA含量较高,APX活性升高,GSSG在高活性GR作用下催化还原为GSH,使得GSH大量积累。因此在匍匐翦股颖ISR抗病反应中,高浓度乙烯促进AsA和GSH的大量积累,它们不仅参与了活性氧的代谢平衡,同时也作为信号分子在ISR抗病反应中起着重要作用。匍匐翦股颖感染褐斑病后,胼胝质主要沉积在厚壁细胞、韧皮部、木质部和表皮组织,其中厚壁细胞胼胝质沉积最多,表皮组织沉积最少。此外,胼胝质沉积面积在不同乙烯信号分子处理间存在差异,但随着处理时间的延长差异不显著。一定浓度的乙烯对ISR反应中胼胝质的沉积具有一定的影响,随着处理时间的延长显著增加,在100 μmol·L^-1ACC处理5 d后,胼胝质沉积总面积仅为9.916 mm²,处理10 d后升至最高,为38.396 mm²,但在病害侵染后期,胼胝质数量减少,在100 μmol·L^-1ACC处理15 d后,胼胝质沉积总面积降至20.052 mm²,反映了乙烯信号分子对胼胝质沉积的影响是一种短期效应,短期内可提高匍匐翦股颖植株抗病性,具有信号分子的短时效特点。研究结果为探清匍匐翦股颖ISR抗病响应中ET信号分子如何调控抗病生理特性提供了理论基础。     

人工改造野生大豆GsDREB2基因对植物耐盐和耐渗透胁迫能力的影响

DREB (dehydration responsive element binding protein)转录因子是一个干旱应答元件的结合蛋白,它能特异结合启动子中含有DRE/CRT顺式元件,激活逆境诱导基因的表达,调控植物对干旱、低温、高盐、高温等胁迫的耐逆性。大量研究表明DREB转录因子在信号传导、作用机理及基因表达方面存在复杂性。为了野生大豆来源GsDREB2基因能更有效地发挥功能,人工突变该基因的负向调节结构域(negative regulatory domain, NRD,140~204),经改造命名为GsDREB2-mNRD。在酵母中比较全长基因(FLDREB2)和GsDREB2-mNRD转录激活和与DRE元件结合的能力,并验证GsDREB2-mNRD核定位情况。分别将FLDREB2和GsDREB2-mNRD转化拟南芥,通过拟南芥幼苗期盐胁迫和渗透胁迫试验,比较GsDREB2-mNRD和FLDREB2在提高植物耐盐和渗透胁迫方面的差异。结果表明,GsDREB2基因内部存在着负向调节结构域(NRD),抑制了GsDREB2转录激活功能和DRE元件结合的特性;经改造的GsDREB2基因依然能定位在细胞核;超量表达GsDREB2-mNRD基因的拟南芥耐盐和渗透胁迫能力明显强于非转基因对照,也高于FLDREB2基因超表达的拟南芥;野生大豆来源的GsDREB2基因NRD结构域的缺失可增强该基因在植物耐盐、渗透胁迫等逆境胁迫下的功能。     

Effect of dasatinib in a xenograft mouse model of canine histiocytic sarcoma and in vitro expression status of its potential target EPHA2

Canine histiocytic sarcoma (HS) is an aggressive and highly metastatic tumor. Previously, the kinase inhibitor dasatinib was shown to have potent growth inhibitory activity against HS cells in vitro, possibly via targeting the EPHA2 receptor. Here, the in vivo effect of dasatinib in HS cells was investigated using a xenograft mouse model. Moreover, the expression status of EPHA2 was examined in six HS cell lines, ranging from insensitive to highly sensitive to dasatinib. In the HS xenograft mouse model, dasatinib significantly suppressed tumor growth, as illustrated by a decrease in mitotic and Ki67 indices and an increase in apoptotic index in tumor tissues. On Western blot analysis, EPHA2 was only weakly detected in all HS cell lines, regardless of sensitivity to dasatinib. Dasatinib likely results in the inhibition of xenograft tumor growth via a mechanism other than targeting EPHA2. The findings of this study suggest that dasatinib is a targeted therapy drug worthy of further exploration for the treatment of canine HS.     

Effects of the sodium‐glucose cotransporter 2 (SGLT2) inhibitor velagliflozin,a new drug with therapeutic potential to treat diabetes in cats

Sodium‐glucose cotransporter 2 (SGLT2) inhibitors are used in the treatment of human diabetics. They increase glucose excretion and correct hyperglycemia. We examined the investigational SGLT2 inhibitor velagliflozin in two groups of six neutered adult obese cats (equal gender distribution). Placebo (Pl) or drug (D; 1 mg/kg) was administered for 35 days. Routine blood examinations, fructosamine, beta‐hydroxybutyrate (BHB), nonesterified fatty acids (NEFA), glucagon, adiponectin, and leptin were measured before and after treatment, also water intake, and urinary electrolytes, glucose, and volume. Indirect calorimetry, an intravenous glucose tolerance test (IVGTT; 0.8 g/kg) and insulin tolerance test (IVITT) were conducted. All cats tolerated treatment well. Significant changes with D included a decrease in the respiratory exchange ratio, an increase in cholesterol, a small increase in albumin, and a rise in BHB and NEFA. Glucose clearance was unaltered, although less insulin was secreted during the IVGTT (p = .056) suggesting improved insulin sensitivity. IVITT was unchanged. Treatment did not affect glucagon, leptin, or adiponectin. Water intake, urine output, urinary glucose excretion, and the glucose/creatinine ratio but not urinary electrolytes were significantly higher post‐D. We conclude that velagliflozin is a promising drug, which increases urinary glucose excretion in cats and could thereby be beneficial for the treatment of hyperglycemia.     

Effects of concentrate‐to‐forage ratios and 2‐methylbutyrate supplementation on ruminal fermentation,bacteria abundance and urinary excretion of purine derivatives in Chinese Simmental steers

This study evaluated the effects of dietary concentrate levels and 2‐methylbutyrate (2MB ) supplementation on performance, ruminal fermentation, bacteria abundance, microbial enzyme activity and urinary excretion of purine derivatives (PD ) in steers. Eight ruminally cannulated Simmental steers (12 months of age; 389 ± 3.7 kg of body weight) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Moderate‐concentrate (400 g/kg diet [MC ]) or high‐concentrate (600 g/kg diet [HC ]) diets were fed with or without 2MB (0 g/day [2MB ?] or 15.0 g/day [2MB +]). Dry matter intake and average daily gain increased, but feed conversion ratio decreased with the HC diet or 2MB supplementation. Ruminal pH decreased, but total volatile fatty acid increased with the HC diet or 2MB supplementation. Molar proportion of acetate and acetate‐to‐propionate ratio decreased with the HC diet, but increased with 2MB supplementation. Propionate molar proportion and ruminal NH ₃‐N content increased with the HC diet, but decreased with 2MB supplementation. Neutral detergent fibre degradability decreased with the HC diet, but increased with 2MB supplementation. Crude protein degradability increased with the HC diet or 2MB supplementation. Abundance of Ruminococcus albus , Ruminococcus flavefaciens , Fibrobacter succinogenes and Bufyrivibrio fibrisolvens as well as activities of carboxymethyl cellulase, cellobiase, xylanase and pectinase decreased with the HC diet, but increased with 2MB supplementation. However, abundance of Prevotella ruminicola and Ruminobacter amylophilus as well as activities of α‐amylase and protease increased with the HC diet or 2MB supplementation. Total PD excretion also increased with the HC diet or 2MB supplementation. The results suggested that growth performance, ruminal fermentation, CP degradability and total PD excretion increased with increasing dietary concentrate level from 40% to 60% or 2MB supplementation. The observed diet × 2MB interaction indicated that supplementation of 2MB was more efficacious for improving growth performance, ruminal fermentation and total PD excretion with promoted ruminal bacteria abundance and enzyme activity in the MC diet than in the HC diet.     

Effects of zinc oxide nanoparticles on the egg quality,immune response,zinc retention,and blood parameters of laying hens in the late phase of production

The objective of this study was to evaluate the effects of dietary supplementation with zinc oxide nanoparticles (ZnO‐NPs) on the performance, egg quality, Zn retention, immunity responses, superoxide dismutase activity (SOD), egg malondialdehyde (MDA) content, and serum parameters in laying hens in the late phase of production. A total of 288 laying hens at 64 weeks of age were randomly assigned to 4 treatments with 6 replicates, and 12 birds within each group. Experimental diets included a corn‐soybean meal‐based diet (without Zn supplementation) and a basal diet supplemented with 80 mg/kg of Zn‐oxide, ZnO‐NPs, and Zn‐methionine. The results indicated that egg production and egg mass were significantly higher in the Zn‐methionine and ZnO‐NPs groups (p < .05). Also, eggshell thickness and shell strength increased in the ZnO‐NPs group as compared with the other groups (p < .05). Moreover, Zn supplementation decreased egg loss (p < .05). There were significant differences among treatments in Zn deposition in tibiotarsus, liver, pancreas, eggs, and excreta (p < .01). Antibody titre, heterophil (%(, and phytohemagglutinin (PHA) were significantly higher in birds fed with Zn‐supplemented diets (p < .05). In treatments supplemented with ZnO‐NPs and Zn‐methionine, the SOD activity in the liver, pancreas, and plasma was greater as compared with the other treatments (p < .05). The MDA content in eggs was significantly reduced in groups supplemented with Zn (p < .01). Moreover, dietary Zn supplementation significantly affected serum total protein, albumin, glucose, alkaline phosphatase activity, carbonic anhydrase activity, and Zn level (p < .05). In conclusion, this study demonstrated that dietary supplementation with ZnO‐NPs can improve the performance of laying hens. Therefore, ZnO‐NPs can enhance zinc absorption in the intestine of aged layers and can be a more suitable source of zinc than regular Zn‐oxide in diets.     

Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.     

小兴安岭天然林不同树种的树高曲线模型

选用10种树高曲线模型作为候选模型,以决定系数(R~2)、残差平方和(S_(se))和均方差(M_(se))作为模型优劣的评价指标,对小兴安岭天然林中的云杉(Picea asperata)、红松(Pinus koraiensis)、水曲柳(Fraxinus mandschurica)、椴树(Tilia tuan)4种树种的树高曲线模型进行优选。结果表明:云杉、红松和椴树的最优模型均为Logistic模型,水曲柳的最优模型为抛物线模型;云杉和红松两种针叶树种,最优模型的优势不明显,水曲柳和椴树两种阔叶树种最优模型的优势较明显;比较R~2的大小,得出较适合小兴安岭云杉和红松两种针叶树种的树高曲线模型(R~20.8),对小兴安岭阔叶树种的适用程度不具有普遍性,对椴树的适用性好(R~2最大达0.94),对水曲柳的适用性较差(R~20.8)。对水曲柳的树高进行分段研究得出:水曲柳在树高低于9 m时,树高曲线模型较适宜(R~20.8),其中抛物线模型为最优模型;树高较高时,没有适用的树高曲线模型。     

三肽囊素对鸡血浆蛋白组分及IL-2与IL-6含量的影响

144只1日龄黄羽肉鸡随机分为4组,第7日龄时第Ⅰ～Ⅳ组分别注射0、0.005、0.010、0.030mg/kg剂量的三肽囊素,在14、35及56日龄时每组各取一个重复,取血浆,醋酸纤维薄膜电泳检测血浆蛋白各组分的比率,放射免疫法检测血浆IL-2与IL-6的含量.结果显示不同剂量的三肽囊素可提高血浆γ球蛋白水平:对血浆...