Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

基于PSR模型的黄河陕西段鱼类增殖放流生态安全评价

基于压力、状态、响应模型(PSR)和层次分析法(AHP),确定17项指标通过数据的标准化处理,指标权重赋值、权重一致性检验、评价等级确定以及评价模型的构建,用生态安全评价黄河陕西段鱼类增殖放流效果,分析生态安全所面临的主要问题。结果表明:黄河陕西段2013年增殖放流生态安全度(ESI)评价等级为Ⅱ级,为良好状态;2014年评价等级为Ⅲ级,处于一般状态;2015年评价等级为Ⅳ级,处于较差状态,属于临界不安全状态以下水平。生态安全形势呈现出逐年下降局势。不安全状态受到影响较大的前3个指标是:黄河径流量变化影响、重要生境保持率和公众资源环境保护意识的影响,3个指标下降值占到下降ESI值的64.93%。其次还受到污水排放达标率、鱼类增殖放流量、政策和管理水平、鱼类生物多样性指数、保护区建设、水质综合污染指数、群落结构等诸多因素影响。研究显示,现阶段增殖放流对黄河生态安全有一定影响,还存在一定提升空间。     

间伐对秦岭南坡锐齿栎林土壤腐殖质及微生物的影响

【目的】以秦岭南坡锐齿栎林为研究对象,探讨间伐对锐齿栎林地土壤腐殖质及土壤微生物的影响,为秦岭林区锐齿栎林抚育经营提供理论依据。【方法】2012年3月,在秦岭南坡陕西省宁东林业局沙沟林场林地阳坡的中下部选择生长状况接近、林分密度基本一致的锐齿栎天然次生林,设置锐齿栎天然次生林样地12块,间伐强度分别为林分蓄积量的0(CK)、5%(T1)、10%(T2)、15%(T3)、20%(T4)和25%(T5),2018年7月采集0～30 cm土层土样,测定土壤腐殖质及其组分(胡敏酸、富啡酸和胡敏素)含量以及土壤微生物(细菌、真菌、放线菌)数量,并分析土壤胡敏酸、富啡酸和胡敏素含量与细菌、真菌、放线菌数量的相关性。【结果】(1)间伐6年后,在0～30 cm土层,各间伐强度处理土壤腐殖质含量平均值均大于CK,且T1处理的土壤腐殖质含量最高,为(60.78±10.53) g/kg。各间伐强度下土壤胡敏酸含量平均值较CK均有明显减少,不同处理土壤胡敏酸含量平均值由大到小依次为CKT4T3T5T2T1。除T3处理外,其余间伐强度处理土壤富啡酸含量平均值均高于CK,富啡酸含量平均值由大到小依次为T2T1T5T4CKT3。与CK相比,除T1处理外,其余各间伐强度处理土壤胡敏素含量平均值均增加,各间伐强度处理土壤胡敏素含量平均值由大到小依次为T2T3T4T5CKT1。(2)不同间伐强度下土壤胡敏酸/腐殖酸含量比值(PQ)由大到小依次为CK15%20%25%10%5%。(3)间伐6年后,在0～30 cm土层,土壤微生物总量随着间伐强度的增加呈先增加后减少的趋势,微生物总量在间伐强度为10%时达到最高;土壤细菌、放线菌数量在间伐强度为5%～10%时较高,真菌数量在间伐强度为15%～20%时较高。(4)土壤细菌、真菌和放线菌数量均与土壤胡敏酸、胡敏素含量呈极显著正相关,土壤真菌数量与富啡酸含量呈显著正相关。【结论】土壤微生物各种群数量可以显著影响土壤腐殖质的积累,合理的抚育间伐可以显著增加土壤腐殖质含量及微生物数量,选择10%～15%的间伐强度作为提高林地土壤腐殖质的参考指标较为合理。     

全基因组选择信号揭示绒山羊绒毛性状相关的候选基因

旨在对辽宁绒山羊和内蒙古绒山羊的基因组选择信号进行筛选。本研究利用Illumina 50K的山羊芯片，对内蒙古绒山羊、辽宁绒山羊和黄淮山羊进行基因型检测，质控后获得50 010个共同SNPs位点。通过群体分化系数Fst和XP-EHH，以黄淮山羊为参考群体分别对内蒙古绒山羊和辽宁绒山羊进行选择信号检测，Fst和XP-EHH值的top 5%作为阈值。结果，利用Fst在绒山羊基因组中筛选到501个候选基因，利用XP-EHH在绒山羊基因组中筛选到145个候选基因。其中有12个SNPs在绒山羊基因组中受到较强选择。通过基因注释筛选到21个候选基因，包括EXOC4、ASIC2、PCDH9、RHBDD1、IRS1和PDE10A等。这些基因主要富集在PI3K-Akt信号通路、蛋白质消化吸收和松弛素信号通路等。研究结果发现，绒山羊在驯化过程中其基因组很多与毛囊相关的基因受到了强烈的选择。这些发现也有助于更好地了解绒山羊的选择进展，为中国绒山羊品种的种质资源保护和利用提供重要参考。     

美国乡村林业发展经验借鉴

在我国经济由高速增长向高质量发展转型的背景下，绿色产业发挥着不可替代的作用。而林业是我国最大的绿色产业，也是构成乡村经济的重要产业部门。文中从林业专业人才、林地管理和林权交易、资金扶持以及乡村产业多元化发展4个方面介绍美国在乡村林业发展的经验，提出促进我国乡村林业发展、推动林业产业扶贫的相关建议，以期为我国林业产业提升经济、社会和生态效益提供参考。     

3种遥感产品检测黄土高原森林变化及差异比较

基于Landsat Vegetation Continuous Fields（LVCF）、MODIS Vegetation Continuous Fields（MVCF）和MODIS Land Cover（MODLC）等遥感产品和森林清查数据（NFI）检测2000-2015年黄土高原森林分布及变化。结果表明:1）2000-2015年黄土高原森林覆盖度升高，基于LVCF和MVCF的全区森林覆盖度均值分别由2000年的7.8%和9.6%增加到2015年的9.7%和13.2%。2）遥感产品估算的森林分布空间格局和面积差异较大，基于LVCF、MVCF和MODLC估算的黄土高原森林面积在2000年分别为726.6×10⁴、604.7×10⁴ hm²和325.1×10⁴ hm²，2015年分别为926.2×10⁴、998.3×10⁴ hm²和400.1×10⁴ hm²。3）遥感产品估算的省级尺度上森林面积与NFI的不一致性因省份和年份而异，LVCF与NFI一致性整体略优于MVCF，MODLC与NFI差异最大且估算的森林面积远低于LVCF、MVCF和NFI。4）遥感产品检测的陕西和山西2省的森林面积增加值与NFI一致性高于宁夏；基于遥感产品估算的宁夏森林面积及增加幅度均远低于NFI。     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

miR-106b-5p靶向KLF4调控山羊肌内前体脂肪细胞分化

旨在明确miR-106b-5p对山羊肌内前体脂肪细胞分化的影响，并确定这种作用是通过靶向KLF4来实现的。本研究利用实时荧光定量PCR （quantitative real-time PCR，qRT-PCR）技术检测miR-106b-5p在山羊肌内前体脂肪细胞分化过程中的表达模式，通过脂质体转染技术将miR-106b-5p mimic和miR-106b-5p inhibitor转入体外培养的山羊肌内前体脂肪细胞，油红O染色法从形态学验证miR-106b-5p对脂肪细胞中脂滴积聚的影响，qRT-PCR检测预测的靶标基因KLF4和脂肪分化标志基因的表达情况，利用双荧光素酶报告系统鉴定miR-106b-5p与KLF4的靶标关系。qRT-PCR结果显示，miR-106b-5p在山羊肌内前体脂肪细胞诱导分化第3天时表达量最高。在山羊肌内脂肪细胞中干扰miR-106b-5p后油红O染色显示脂滴聚积减少，过表达miR-106b-5p后脂滴聚积增加。在山羊肌内前体脂肪细胞中转染miR-106b-5p inhibitor后PPARγ表达量显著降低（P<0.05），而KLF4表达量极显著升高（P<0.01）；转染miR-106b-5p mimic后LPL和PPARγ表达量极显著升高（P<0.01）。荧光素酶活性试验结果显示，过表达miR-106b-5p可显著抑制KLF4荧光活性。miR-106b-5p通过靶向并负调节KLF4的表达促进山羊肌内脂肪细胞分化。     

Effects of different active constituents of Gynostemma pentaphyllum on mitochondrial energy metabolism-related proteins in endothelial cells induced by ox-LDL

AIM To investigate the effects of different components of Gynostemma pentaphyllum ［gypenosides (Gps), gypenoside XLIX (GpXLIX) and ginsenoside Rb3 (GRb3)］ on mitochondrial energy metabolism-related proteins in endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). METHODS EA.hy926 cells were divided into control group, model group, Gps group, GpXLIX group and GRb3 group. The cells in control group were cultured only in DMEM complete medium. The cells in model group were treated with 100 mg/L ox-LDL for 48 h. The cells in Gps group, GpXLIX group and GRb3 group were treated with 100 mg/L ox-LDL for 24 h, and then treated with Gps, GpXLIX and GRb3 at 100 mg/L for another 24 h, respectively. The ATP content in each group was detected by ELISA. The expression levels of mitochondrial energy metabolism-related proteins, cytochrome C oxidase subunit 5a （Cox5a）, NADH:ubiquinone oxidoreductase core subunit S1 （Ndufs1）, ATP synthase F1 subunit alpha (ATP5a) and cytochrome C (Cyt C）, were determined by Wes automatic Western blot quantitative analysis system and Western blot. RESULTS Compared with control group, the ATP content in model group was decreased (P<0.01). After drug intervention, the ATP content increased to different degrees in Gps group, GpXLIX group and GRb3 group (P<0.01). The results of Wes automatic Western blot quantitative analysis system were consistent with those of Western blot. These results showed that compared with control group, the protein expression of Cox5a, Ndufs1 and ATP5a in model group was decreased, and the protein expression of Cyt C was increased (P<0.01). After intervention, the protein expression of Cox5a, Ndufs1 and ATP5a was increased and the protein expression of Cyt C was decreased in Gps group, GpXLIX group and GRb3 group (P<0.05 or P<0.01). Among them, the effect of Gps on the protein expression of Cox5a, Ndufs1 and Cyt C was significantly stronger than those of the 2 monomer components, and the effect of GRb3 was found to be superior in the 2 monomer components. The effect of GpXLIX on ATP5a protein was superior to the other 2 components. CONCLUSION Gynostemma total saponins and related active ingredients protect ox-LDL-induced endothelial cells by affecting mitochondrial energy metabolism-related proteins, thereby preventing and treating atherosclerosis.