Changes of Ca2＋ activated potassium channels and cellular-proliferation in autogenous vein grafts

AIM: To investigate changes of Ca^2＋ activated potassium channels (K_Ca) in autogenous vein grafts. METHODS: The contraction of venous ring was measured by means of perfusion in vitro. The intimal proliferation and proliferation of cultured smooth muscle cells (vascular smooth muscle cells, VSMCs) were observed by the means of computerised image analysis and MTT method, respectively. Furthermore, whole cell mode of patch clamp was used to record K_Ca of VSMCs isolated from autogenous vein grafts. RESULTS: 1 week after transplantation there were no significant differences of contraction and intimal relative thickness between autogenous vein grafts and control. Contraction and intimal relative thickness of autogenous vein graft were significantly increased 2 weeks after transplantation (P＜0.05, n=8 vs control), and they were more enhanced 4 weeks after vein transplantation (P＜0.01, n=8 vs control). TEA (blocker of Ca^2＋ activated potassium channels) increased MTT A₄₉₀ value of VSMCs from femoral vein in a dose-dependent manner (P＜0.05, n=8). K_Ca current density was significantly attenuated in VSMCs from autogenous vein grafts 1-4 weeks after transplantation (P＜0.05, n=5). CONCLUSION: K_Ca was inhibited in autogenous vein graft, which accounted for vasospasm and intimal proliferation.     

NO介导的Ca2+信号对渗透胁迫下紫花苜蓿幼苗光合特征及抗性的影响

以紫花苜蓿幼苗为材料,用聚乙二醇(PEG-6000)作为渗透介质人工模拟干旱条件,外源喷施NO供体硝普钠(SNP)、钙信号试剂CaCl₂、NO抑制剂亚甲基蓝(MB)和Ca²⁺通道阻断剂LaCl₃,对紫花苜蓿幼苗光合特征、抗氧化酶活性及过氧化物酶(POD)同工酶图谱进行研究,探讨了渗透胁迫下NO介导的Ca²⁺信号对紫花苜蓿幼苗光合作用及抗氧化能力的影响。结果表明:在渗透胁迫条件下,施加SNP、CaCl₂均能够有效缓解叶片叶绿素a、类胡萝卜素及总叶绿素含量降低,提高叶片净光合速率(P_n)、气孔导度(G_s)及气孔限制值(L_s),而对胞间CO₂浓度(C_i)没有缓解作用。SNP、CaCl₂及SNP+CaCl₂处理提高了幼苗叶片中抗氧化酶活性和脯氨酸含量,降低了丙二醛(MDA)含量。其中共处理时效果最为显著,第4天 SOD、POD、CAT活性较PEG处理升高了39.29%、30.41%和56.24%,脯氨酸含量增加了45.59%,MDA含量降低了45.59%。POD同工酶图谱在第4天时酶谱带数最多,POD活性最强,且SNP+CaCl₂共处理下出现新酶带。而添加外源NO的同时添加Ca²⁺通道阻断剂LaCl₃,紫花苜蓿幼苗光合速率、抗氧化酶活性及脯氨酸含量均降低,丙二醛含量增加,添加Ca²⁺信号的同时施加NO抑制剂MB也具有相同的作用,说明Ca²⁺信号参与NO信号转导过程并相互作用共同调节渗透胁迫下紫花苜蓿幼苗的生理应答响应。     

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.     

基于紫花苜蓿混播草地的全草型肉羊放牧育肥模式案例研究

本研究旨在探讨紫花苜蓿（Medicago sativa L.）混播草地放牧利用的生产潜力和集成全草型集约化肉羊放牧育肥技术模式。2017-2018年，在位于河北省廊坊市的中国农业科学院国际农业高新技术产业园内开展了2年羔羊育肥的划区轮牧试验，研究了紫花苜蓿混播草地的地上生物量、育肥羊增重以及羊肉品质的变化规律。结果表明：混播草地地上生物量超过11.2 tDM·hm^-2，5月份紫花苜蓿混播草地的粗蛋白为20.92%，每公顷紫花苜蓿混播草地可满足51只育肥羊、放牧育肥150天，每只增重30 kg所需的干物质、代谢能和粗蛋白三个方面的全部需求。与此同时，紫花苜蓿混播草地还可以显著增加羊肉中欧米伽3型多不饱和脂肪酸的含量，n-6/n-3比例为2.72，羊肉健康品质显著提高。以紫花苜蓿混播草地为基础的全草型肉羊放牧育肥技术具有广阔的应用前景，对我国苜蓿产业以及草地畜牧业发展具有重要意义。     

Effect of the particle size on the ammonia removal rate and the bacterial community composition of bioflocs

The total ammonia nitrogen (TAN) removal efficiency and bacterial community composition of bioflocs with <50-μm particle size, > 50-μm particle size and un-sieved bioflocs were investigated in the current study. The initial ratio of dissolved organic carbon to TAN (DOC/TAN) in the three groups were about 14:1. No significant difference was found in the removal rate of TAN, average concentrations of TAN and nitrite nitrogen among the three groups (P > 0.05). The C/N (w/w) ratio of the > 50-μm bioflocs was significantly higher than those of the other groups. No significant differences were found in the crude protein content in the bioflocs among the three groups. The development of the bacterial community compositions of the bioflocs was analyzed by Illumina MiSeq sequencing analyses. Most OTUs were shared among the three groups at all the sampled time points. With the increase in the relative abundance of phylum Firmicutes, that of phylum Proteobacteria, Chorolexi, and Bacteroidetes decreased in all the three groups. The phylum Firmicutes and genus Bacillus were predominant in all the sampled time points. At the end of the experiment, genus Bacillus accounted for 81% in the < 50-μm group, 82% in the > 50-μm group, and 75% in the un-sieved group.     

Sugar beet rotation effects on soil organic matter and calculated humus balance in Central Germany

In order to quantify the influence of land use systems on the level of soil organic matter (SOM) to develop recommendations, long-term field studies are essential. Based on a crop rotation experiment which commenced in 1970, this paper investigated the impact of crop rotations involving increased proportions of sugar beet on SOM content. To this end, soil samples were taken in 2010 and 2012 from the following crop rotation sequences: sugar beet–sugar beet–winter wheat–winter wheat (SB–SB–WW–WW = 50%), sugar beet–sugar beet–sugar beet–winter wheat (SB–SB–SB–WW = 75%), sugar beet–grain maize (SB–GM = 50%) and sugar beet-monoculture (SB = 100%); these were analysed in terms of total organic carbon (TOC) and microbial biomass carbon (MBC) content, MBC/TOC ratio and the TOC stocks per hectare. In addition, humus balances were created (using the software REPRO, reference period 12 years) in order to calculate how well the soil was supplied with organic matter. In the field experiment, harvest by-products (WW and GM straw as well as SB leaves) were removed. After 41 years, no statistically significant differences were measured between the crop rotations for the parameters TOC, MBC, MBC/TOC ratio and the TOC stock per hectare. However, the calculated humus balance was significantly affected by the crop rotation. The calculated humus balance became increasingly negative in the order SB–SB–WW–WW, SB–SB–SB–WW, SB monoculture and SB–GM, and correlated with the soil parameters. The calculated humus balances for the reference period did not reflect the actual demand for organic matter by the crop rotations, but instead overestimated it.     

Dietary mannitol increased the absorption of calcium and magnesium in rats

The effect of mannitol on bone‐related mineral absorption and retention and the mechanism was investigated in this study. Fourteen 8‐week‐old male Wistar rats in experiment 1 and same number and age cecectomized Wistar male rats in experiment 2 were divided into two subgroups of seven animals, respectively, fed diets containing 0 or 4% mannitol for 28 days. Mineral balance tests were determined twice during days 8–12 and days 22–26, and the rats were slaughtered on day 28 both in experiment 1 and experiment 2. The whole caecum and colon were collected with the content to analyse tissue weight, content weight, content's pH and moisture, organic acids' concentration and mineral levels. In experiment 1, Ca absorption and retention and Mg absorption were significantly increased by mannitol feeding during days 8–12. Caecal total weight, tissue weight and content weight were increased, the pH of caecum and colon was reduced, and the concentrations of caecal short‐chain fatty acids (SCFAs) were modified by mannitol feeding. In experiment 2, during days 8–12 and days 22–26, Ca absorption and retention were significantly lowered by mannitol feeding in cecectomized rats; however, mannitol feeding decreased Mg absorption during days 8–12, but did not impact Mg retention. Colonic total weight, tissue weight and content weight were significantly increased, and colonic pH was reduced by mannitol feeding. In conclusion, dietary mannitol increased the absorption of Ca and Mg and the caecum markedly contributed to this promoting effect of mannitol.     

Binding Sites for Ca2+-Channel Effectors and Ryanodine in Periplaneta americana—Possible Targets for New Insecticides

The calcium channel and the ‘calcium release channel’ of muscle membrane of the cockroach Periplaneta americana have been characterized. Biological assays with calcium channel blockers and ryanodine on different insects and acari revealed pronounced insecticidal effects with ryanodine, but not with calcium channel blockers, at concentrations between 0·1 and 300 μg ml⁻¹. Skeletal muscle membranes derived either from the tubular network or from the sarcoplasmatic reticulum of P. americana were characterized with respect to the binding of the dihydropyridine (DHP) [³H]isradipine (PN 200-110), the phenyl-alkylamine [³H]verapamil and the alkaloid [³H]ryanodine. Preliminary binding studies with the benzothiazepine [³H]diltiazem suggest a low-affinity binding site with a IC₅₀ value of 3·3 μM . All binding sites tested were sensitive to treatment with proteinase K. Optimal conditions for binding of the radioligand ryanodine revealed the highest specific binding at pH 8 and at calcium chloride concentrations between 100 and 500 μM . EGTA at 10 μM abolished 95% of the ryanodine binding. Binding studies with calcium channel binding sites revealed a pronounced effect of low Ca²⁺ concentrations on specific isradipine binding, whereas verapamil and diltiazem binding were only reduced by the presence of 200 μM EGTA. With respect to high Ca²⁺ concentrations, specific binding of diltiazem, isradipine and verapamil was reduced by 73, 40 and 20%, respectively, at 5 mM Ca²⁺. Radioligand binding experiments showed high-affinity binding sites for ryanodine and isradipine. K_D values of 0·95 nM (B_max=550 fmol mg⁻¹ protein) and 0·75 nM (B_max=213 fmol mg⁻¹ protein) were determined respectively. A lower-affinity binding site was identified in binding studies with verapamil (K_D=7·4 nM and B_max=27 fmol mg⁻¹ protein). [³H]isradipine displacement studies with several dihydropyridines revealed the following ranking of affinity: nitrendipine>isradipine>Bay K8664≪nicardipine. Displacement of [³H]verapamil binding by effectors of the phenylalkylamine binding site showed that bepridil and S(-)verapamil had the highest affinities of the compounds tested followed by (±)verapamil, nor-methylverapamil and R(+)verapamil.     

灰斑病菌对大豆叶片总多酚和总黄酮的诱导研究

利用大豆灰斑病菌不同致病力的生理小种接种抗感不同的大豆品种,结果表明:大豆叶片内总黄酮类物质的含量与品种的抗病性呈明显正相关;感病品种叶片内总多酚含量变化显著降低,抗病品种总体变化不大.接种致病力强的生理小种能够大幅度提高抗病品种体内总黄酮含量,这一结果可以作为筛选和鉴定抗病品种的一种生化指标;大豆灰斑病菌弱毒菌株能够诱导感病品种体内总黄酮含量提高,进而提高大豆的抗病性,这一结果为大豆灰斑病的生物防治提供了依据.