Coupling changes of soil functional gene abundances and extracellular enzyme activities across the diagnostic horizons of agricultural Isohumosols

Soil functional microbial taxa and extracellular enzymes are involved in a variety of biogeochemical cycling processes. Although many studies have revealed the vertical change patterns of microbial communities along soil profile, the general understanding of the coupling changes in the functional gene abundances (FGAs) and extracellular enzyme activities (EEAs) in soil profiles is still limited, which hinders us from revealing soil ecosystem processes. Herein, we comparatively investigated the FGAs and EEAs in the diagnostic A, B, and C horizons of soil profiles obtained from two suborders of Isohumosols (Mollisols), Ustic and Udic Isohumosols, in Northeast China based on quantitative real-time polymerase chain reaction and standard fluorometric techniques, respectively. The distribution patterns of both FGAs and EEAs significantly distinguished by the two soil suborders and were also separated from A to C horizon. Additionally, the variations of EEAs and FGAs were greater in Udic Isohumosols compared to Ustic Isohumosols along soil profiles, and greater changes were observed in C horizon than in A horizon. Both FGAs and EEAs correspondently decreased along the soil profiles. However, when normalized by soil organic carbon, the specific EEAs significantly increased in deep soil horizons, suggesting that microorganisms will input more resources to the production of enzymes to ensure microbial nutrient requirements under resource scarcity. More importantly, we revealed that soil microbial nutrient demands were limited by carbon (C) and phosphorus (P), and the C and P limitations significantly increased along soil profiles with a greater C limitation observed in Ustic Isohumosols than in Udic Isohumosols. Overall, our findings provided solid evidence showing the links between FGAs, EEAs, and microbial nutrient limitations, which would be helpful for a better understanding of the ecosystem processes in soil profiles.     

Resistance and resilience of soil biological indicators: A case study with multi-walled carbon nanotube

Soil ecosystem is experiencing stresses due to climate change, and soil inhabitants try to demonstrate their inherent resistance and resilience against those stresses. Application of nanomaterials as agricultural inputs could bring shifts in resistance and resilience patterns of soil microbes and associated enzymes, especially under short-term heat stress. With this background, the impacts of multi-walled carbon nanotube (MWCNT) on the resistance and resilience of soil biological indicators were evaluated. An incubation experiment was conducted with varied MWCNT concentrations (0, 50, 100, 250, and 500 mg kg^-1 soil) for 90 d after 24-h heat stress at 48 ±2 ℃ to assess the impacts of MWCNT on soil enzyme activities and microbial populations vis-à-vis their resistance and resilience indices under short-term exposure to heat stress. Enzyme activities were reduced after exposure to heat stress. Resistance indices of enzyme activities were enhanced by MWCNT application on day 1 after heat stress, whereas there was no recovery of enzyme activities after 90-d incubation. Like soil enzyme activities, resistance index values of soil microbial populations followed the similar trend and were improved by MWCNT application. Multi-walled carbon nanotube has the potential to improve resistance indices of soil enzyme activities and microbial populations under heat stress, although they could not recover to their original state during periodical incubation after heat stress. This study helps to understand the relative changes of biological indicators under MWCNT and their ability to withstand heat stress.     

饲粮能量和蛋白质水平对滩羊小肠中小肽和氨基酸转运载体mRNA表达量的影响

本试验旨在研究饲粮能量和蛋白质水平对滩羊小肠中小肽和氨基酸转运载体mRNA表达量的影响。选取112只健康、体重相近的滩羊,随机分成4组,每组4个重复,每个重复7只羊。标准水平的饲粮能量和蛋白质水平参考《肉羊饲养标准》(NY/T 816—2004),各组试验滩羊分别饲喂不同能量和蛋白质水平饲粮:0.84×标准水平(Ⅰ组)、0.96×标准水平(Ⅱ组)、1.08×标准水平(Ⅲ组)和1.20×标准水平(Ⅳ组)。试验根据羊体重分2个阶段:29~35 kg和36~40 kg。于每个阶段末,每个重复屠宰1只试验羊,取其小肠组织样,运用实时荧光定量PCR技术,研究小肽转运载体1(Pep T1)、y+型氨基酸转运载体1(CAT1)、兴奋性氨基酸转运载体3(EAAT3)mRNA表达量的变化。结果表明:1)在29~35 kg阶段末,小肠中Pep T1 mRNA的表达量随着饲粮能量和蛋白质水平的提高呈先下降再上升的趋势,Ⅱ组显著低于其他3组(P0.05);Ⅳ组小肠中CAT1 mRNA的表达量显著高于其他3组(P0.05);Ⅲ组小肠中EAAT3mRNA的表达量显著高于其他3组(P0.05)。2)在36~40 kg阶段末,Ⅱ组小肠中Pep T1mRNA的表达量显著高于其他3组(P0.05);Ⅱ组小肠中CAT1 mRNA的表达量显著高于Ⅲ组(P0.05);小肠中EAAT3 mRNA的表达量随着饲粮能量和蛋白质水平的提高呈上升趋势,Ⅲ组和Ⅳ组小肠中EAAT3 mRNA的表达量显著高于Ⅰ组和Ⅱ组(P0.05)。由此可见,饲粮能量和蛋白质水平会影响滩羊小肠中Pep T1、CAT1、EAAT3 mRNA的表达量,使机体对小肽和氨基酸的吸收利用率随之改变,以适应滩羊的生长发育。     

低温处理对紫苏生理指标的影响

[目的]研究低温处理对紫苏生理指标的影响,确定造成紫苏叶片低温伤害的时间。[方法]将紫苏叶分别低温处理0、12、24、36、48和60 h,并测定其叶绿素、可溶性蛋白质及可溶性糖的含量,并考察2～5℃低温对紫苏叶片生理指标的影响,确定紫苏叶片低温伤害的时间。[结果]低温处理48 h能明显减少叶片中叶绿素的含量,而且低温处理60 h的叶绿素含量减少非常明显;低温处理48 h能明显减少叶片中可溶性蛋白质和可溶性糖的含量。[结论]低温处理超过48 h,紫苏就会受到伤害;低温处理时间达到60 h时,紫苏受到的伤害程度加重。     

Atorvastatin inhibits IL-1β and IL-18 releases via lowering the expression of NLRP1 inflammasome

AIM: To explore the role of nucleotide-binding oligomerization domain-like receptor protein 1 (NLRP1) inflammasome in atorvastatin-induced reduction of interleukin-1β (IL-1β) and interleukin-18 (IL-18) releases from the THP-1 macrophages. METHODS: Lipopolysaccharide (LPS, 10 μg/L) was used to trigger the secretion of IL-1β and IL-18 in the THP-1 macrophages. The cells were incubated with different concentrations of atorvastatin (1, 10 and 20 μmol/L) for 24 h, or treated with 10 μmol/L atorvastatin for different time (12 h, 24 h and 48 h). NLRP1 siRNA was transfected into the THP-1 cells. The mRNA expression of NLRP1 inflammasome was detected by RT-PCR. The protein expression of NLRP1 inflammasome was determined by Western blot. The secretion of proinflammatory cytokines IL-1β and IL-18 was quantified by ELISA. RESULTS: Atorvastatin inhibited the mRNA and protein expression of NLRP1 inflammasome in the THP-1 macrophages in a dose- and time-dependent manner. Transfection of NLRP1 siRNA significantly decreased the protein expression of NLRP1 and promoted the suppressive effect of atorvastatin on IL-1β and IL-18 secretion in the THP-1 macrophages. CONCLUSION: Atorvastatin inhibits the production of IL-1β and IL-18 in the macrophages through decreasing NLRP1 inflammasome expression, possibly contributing to the anti-inflammatory effect of atorvastatin on atherosclerosis.     

含蛋氨酸二肽影响奶牛乳腺上皮细胞乳蛋白合成相关基因表达

本试验旨在研究含蛋氨酸(Met)二肽对奶牛乳腺上皮细胞(BMECs)内乳蛋白合成相关基因表达的影响。试验分3部分,均采用单因子完全随机试验设计,Met的添加浓度及培养时间分别为60μg/m L(0.402 mmol/L)、48 h。第1部分,培养液添加8种含Met二肽[蛋氨酸-蛋氨酸(P-Met-Met)、蛋氨酸-赖氨酸(P-Met-Lys)、蛋氨酸-色氨酸(P-Met-Trp)、蛋氨酸-苯丙氨酸(P-Met-Phe)、蛋氨酸-苏氨酸(P-Met-Thr)、蛋氨酸-异亮氨酸(P-Met-Ile)、蛋氨酸-亮氨酸(P-Met-Leu)、蛋氨酸-缬氨酸(P-Met-Val)],以不添加二肽为对照,测定BMECs乳蛋白合成相关基因(αs1-酪蛋白、β-酪蛋白、κ-酪蛋白、β-乳球蛋白、Ⅱ型小肽转运载体和氨肽酶氮)的表达量;第2部分,培养液添加8种与上述二肽对应的游离氨基酸(F-Met-Met、F-Met-Lys、F-MetTrp、F-M et-Phe、F-M et-Thr、F-M et-Ile、F-M et-Leu、F-M et-Val),以不添加游离氨基酸为对照,测定BM ECs乳蛋白合成相关基因的表达量;第3部分,用二肽等物质的量替代相应游离氨基酸,测定BMECs乳蛋白合成相关基因的表达量以及细胞内外氨肽酶含量。结果表明:P-Met-Met和P-M et-Lys组较对照组和其他二肽组上调了αs1-酪蛋白和β-酪蛋白基因的表达量,且P-M et-M et组优于P-Met-Lys组。F-Met-Met和F-Met-Lys组较对照组和其他游离氨基酸组显著提高了αs1-酪蛋白基因的表达量(P0.05)。除P-Met-Val和P-Met-Leu组外,其他二肽替代游离氨基酸后均不同程度地提高了乳蛋白和Ⅱ型小肽转运载体基因的表达量,其中P-Met-Met表现出较好的促进效果。总之,含Met二肽等量替代对应的游离氨基酸能够促进乳蛋白基因的表达,其中尤以P-Met-Met的效果最好。     

Specific polyphenol oxidase activity of red clover (Trifolium pratense) and its relation with forage quality in field experiments

Abstract

In feeding studies, red clover (RC) influenced positively the N utilization by ruminants. A relationship between polyphenol oxidase (PPO) activity and forage quality has not been established. Our objective was to investigate seasonal, site, genotype, and management effects on specific PPO activity in RC in three experiments under field conditions, and relate the activity to forage-quality parameters. In Experiment 1, six RC genotypes at two study sites were submitted to a 3-cut system. Specific PPO activity, forage quality, and vegetation stage were determined. PPO activity varied between harvest and study sites, with genotypes differing up to 3-fold in PPO activity within harvests. The specific PPO activity, forage quality, and vegetation stage in RC subjected to 5-cut system and grazing (Experiment 2) were determined. Additionally, in Experiment 3, cutting frequency in RC swards including mechanical stress (rolling) was investigated. The induction of PPO activity in RC by grazing or mechanical stress (Experiments 2 and 3) increased the activity up to 2.5-fold compared with RC at similar vegetation stage submitted to the 5-cut system. Mechanical stress induced by grazing or rolling, and seasonal differences, seem to have a larger influence on specific PPO activity than does the genotype effect observed in Experiment 1. For forage quality, an increased specific PPO activity explained 29–46% of the reduction in protein fraction ‘A’ content (non-protein N) in the cutting systems in Experiments 2 and 3. Other CP fractions achieved a lower relation. Furthermore, the precipitation-to-temperature ratio preceeding a harvest explained 63% of the variation in the specific PPO activity. In conclusion, the PPO activity in RC is induced by grazing and rolling. Whereas weather conditions preceeding a harvest showed a large influence, genotype influence had only minor relevance. These results may have implications for regional harvest management towards efficient N utilization by ruminants.     

人呼吸道合胞病毒的研究与防治进展

人呼吸道合胞病毒是全世界婴幼儿下呼吸道感染最常见的病原体,导致每年超过300万人住院和20万以上的人死亡,给全球带来了非常严重的经济和社会负担。目前,尚不存在防治RSV的疫苗,也没有特异性强、安全性高的抗RSV药物。对HRSV的结构、流行病学特征以及疾病模型构建方面的研究进展进行综述,并对已有的检测和治疗方法进行总结,旨在为今后开发抗HRSV的药物提供全面的参考信息。     

应用VITEK2全自动微生物鉴定系统检测养殖用水中的大肠杆菌

采用国家相关的检验养殖用水标准方法、API 20E微生物生化试验、VITEK2全自动微生物鉴定系统，对养殖用水样品进行了大肠杆菌的检测方法比较和鉴定。VITEK2鉴定系统自动打印的实验报告和API 20E微生物生化试验的判读比对结果显示：3个参试样品分别为阴沟肠杆菌、产气肠杆菌和大肠埃希菌，VITEK2与API 20E两种方法的鉴定结果符合率100%。从实验可知，利用VITEK2全自动微生物鉴定系统，对被污染的养殖用水中细菌源的鉴定速度快，方法简便，结果准确、实用。此方法也可用于各种污染环境及食品污染源的     

产单核细胞李斯特菌P60蛋白的可溶性表达

以食品中分离到的L.mXFL0605株为试验菌株,PCR扩增得到iap全基因。将iap连接到pET-28a(+)载体进行原核表达,分别采用培养温度为25℃或37℃,IPTG浓度为5μg/mL或10μg/mL优化组合进行诱导。结果显示,当采用37℃培养和10μg/mLIPTG诱导时可以获得部分可溶性表达的p60,重组p60蛋白分子质量约60 ku。