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.     

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.     

不同温度下圆叶决明降解过程红壤可溶性氮及氮水解酶动态变化研究

为了探明圆叶决明（Chamaecrista rotundifolia）降解过程果园红壤供氮水平的变化规律，本研究采用模拟培养试验，研究15℃和25℃培养果园红壤硝态氮、铵态氮、可溶性总氮、可溶性有机氮含量的变化及脲酶、蛋白酶、天冬酰胺酶活性的变化。结果表明，圆叶决明降解过程果园红壤4种可溶性氮含量均显著提高，且在培养140 d达到最大值，25℃培养的效果更佳。其中，可溶性总氮、可溶性有机氮和硝态氮含量随时间的动态变化可用三次曲线方程来拟合。圆叶决明降解过程还能显著提高脲酶、蛋白酶和天冬酰胺酶的活性，其中蛋白酶活性和脲酶活性的变化可用指数方程和三次曲线方程来拟合。本研究认为圆叶决明降解过程可提高果园红壤可溶性氮含量，因此可通过翻压圆叶决明提高土壤的供氮水平。     

Response of labile organic C and N pools to plastic film removal from semiarid farmland soil

To examine the effects of plastic film removal on grain yield and soil organic matter (SOM), a spring maize (Zea may L.) field experiment was conducted for 5 yr at Changwu Agricultural and Ecological Experimental Station of Northwest China. Compared with traditional plastic film mulching during entire growing stages (FM), plastic film removal at the silking stage (RM) resulted in a 6.3% higher average maize yield. Under the RM treatment, soil organic carbon and total nitrogen significantly increased after the 5‐yr cultivation in the 0‐ to 20‐cm layer. Significant increases in extractable organic C (EOC), KMnO₄‐oxidizable C (KMnO₄‐C) and C management index (CMI) in the 0‐ to 20‐cm layer, and light fraction organic C and EOC in the 20‐ to 40‐cm layer were observed in response to plastic film removal after the 1‐yr treatment; the responses were more significant after 5 yr. Under the RM treatment, significant increases in microbial biomass C, light fraction organic N, extractable organic N, KMnO₄‐C and CMI were also observed after five years in the 20‐ to 40‐cm layer. Moreover, KMnO₄‐C and EOC were much more sensitive than other labile SOM fractions to the application of RM, even after only 1 yr of cultivation. Therefore, compared with mulching for the whole growing season, plastic film removal at the maize silking stage is an effective option for increasing yields and enhancing SOM concentration and soil sustainability in the regions with semiarid monsoon climates that have sufficient rainfall during maize reproductive stages.     

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...     

外源氮添加对湿地土壤N_2O排放量的影响

为搞清湿地土壤驱动N2O排放的关键氮源类型,有效减少湿地N2O的排放,本文通过室内控制温湿度,用气相色谱法分析不同外源氮素对湿地N2O排放的影响。结果表明:外加氮源组总是高于对照组N2O排放量(4.4 mg·m-3)。在设定的剂量范围内,单独添加尿素或尿素与硝酸铵1∶1配合时N2O排放量呈现先增后减的单峰分布趋势,峰值分别为10.6 mg·m-3和229.0 mg·m-3;单独添加硝酸铵时N2O排放量(32.6~111.0 mg·m-3)随着氮素添加量增加呈现持续上升趋势。单独添加尿素或硝酸铵、尿素与硝酸铵1∶1配合均促进N2O的排放,但硝酸铵尿素混合添加对N2O排放量的贡献单独添加硝酸铵单独添加尿素。这为预测内蒙古高原区农牧交错带湿地氮素输入可能带来的温室效应和有效减排提供科学依据。