Effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE-/- mice

AIM To observe the effect of tanshinone ⅡA on liver lipid deposition and ferroptosis-related protein expression in ApoE^-/- mice. METHODS Thirty-two ApoE^-/- mice were randomly divided into model group, high-dose (60 mg/kg) tanshinone ⅡA group, low-dose (30 mg/kg) tanshinone ⅡA group and simvastatin group, and C57BL/6J mice (n=8) were used as normal control group. The mice in normal control group were given the basic feeding, while the others were given high-fat diet. The mice in tanshinone ⅡA groups and simvastatin group were given corresponding drugs. The mice in normal control group and model group were intraperitoneally injected with equal volume of saline. Eight weeks later, the serum levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were tested by automatic biochemistry analyzer. The liver tissues were stained with HE and oil red O. The contents of reactive oxygen species (ROS) and glutathione (GSH) in liver tissues of the mice were measured by commercially available kits. The liver glutathione peroxidase 4 (GPX4) and p53 were detected by immunohistochemical method. The protein and mRNA expression levels of ferroptosis-related factors GPX4, xCT/SLC7A11, p53 and ferritin heavy chain 1 (FTH1) were determined by Wes automatic Western blot quantitative analysis system and RT-qPCR. RESULTS Compared with normal control group, the serum levels of TC, TG and LDL-C in model group were increased significantly (P<0.05 or P<0.01), and HDL-C did not change significantly. The fat vacuoles were clearly visible in liver tissue. The content of ROS in liver tissue was increased significantly,and GSH was decreased significantly (P<0.01). The mRNA and protein expression levels of p53 were increased significantly, and GPX4, xCT/SLC7A11 and FTH1 were decreased significantly (P<0.05 or P<0.01). Compared with model group, tanshinone ⅡA significantly decreased the serum levels of TC, TG and LDL-C (P<0.05 or P<0.01), and HDL-C did not change significantly. High-dose and low-dose tanshinoneⅡA also significantly decreased the degree of steatosis, and the size of lipid droplets. The content of ROS in liver tissues was decreased significantly, and GSH was increased significantly (P<0.01). The mRNA and protein expression levels of GPX4, xCT/SLC7A11 and FTH1 were increased significantly, and p53 were decreased significantly (P<0.05 or P<0.01). CONCLUSION Tanshinone ⅡA reduces liver lipid deposition and lipid peroxidation damage in ApoE^-/- mice, which may be related to the intervention of ferroptosis-related proteins in the liver cells.     

Impact of native form oat β-glucan on starch digestion and postprandial glycemia

Whole grain oats, as a rich source of β-glucan, have been shown beneficial to glycemic control. In the current study, the impact of native form β-glucan in oat grains (NFO-glucan) on starch digestion and postprandial glycemia was investigated. The dry-milling prepared NFO-glucan sample was enriched with native form β-glucan (15.6%), and an in vitro starch digestion assay of NFO-glucan (0.5% starch equivalent) showed a significant decrease of starch digestion rate compared to oat starch (0.5%, w/v). However, pretreatment by either β-glucanase or pepsin significantly increased the starch digestion. Consistently, an in vivo examination on the postprandial glycemia of the cooked NFO-glucan sample using a mouse model displayed a significant decrease of postprandial glycemia compared to gelatinized oat starch. Further experiment on the pasting property of NFO-glucan sample by a rapid visco-analyser demonstrated both β-glucan and protein affected its viscosity profiles. Scanning Electronic Microscope (SEM) observation revealed a network-like native structure of β-glucan that might encapsulate protein and starch to reduce the enzyme accessibility and so the digestion of starch. Novel food processing technologies to maintain the native form of β-glucan in oat grains might be a better way to modulate the postprandial glycemia of oat-based whole grain foods.     

Substitution of chemical fertilizer by Chinese milk vetch improves the sustainability of yield and accumulation of soil organic carbon in a double-rice cropping system

The double-rice cropping system is a very important intensive cropping system for food security in China. There have been few studies of the sustainability of yield and accumulation of soil organic carbon(SOC) in the double-rice cropping system following a partial substitution of chemical fertilizer by Chinese milk vetch(Mv). We conducted a 10-year(2008–2017) field experiment in Nan County, South-Central China, to examine the double-rice productivity and SOC accumulation in a paddy soil in response to different fertilization levels and Mv application(22.5 Mg ha~(–1)). Fertilizer and Mv were applied both individually and in combination(sole chemical fertilizers, Mv plus 100, 80, 60, 40, and 0% of the recommended dose of chemical fertilizers, labeled as F100, MF100, MF80, MF60, MF40, and MF0, respectively). It was found that the grain yields of double-rice crop in treatments receiving Mv were reduced when the dose of chemical fertilizer was reduced, while the change in SOC stock displayed a double peak curve. The MF100 produced the highest double-rice yield and SOC stock, with the value higher by 13.5 and 26.8% than that in the F100. However, the grain yields increased in the MF80(by 8.4% compared to the F100), while the SOC stock only increased by 8.4%. Analogous to the change of grain yield, the sustainable yield index(SYI) of double rice were improved significantly in the MF100 and MF80 compared to the F100, while there was a slight increase in the MF60 and MF40. After a certain amount of Mv input(22.5 Mg ha~(–1)), the carbon sequestration rate was affected by the nutrient input due to the stimulation of microbial biomass. Compared with the MF0, the MF100 and MF40 resulted in a dramatically higher carbon sequestration rate(with the value higher by 71.6 and 70.1%),whereas the MF80 induced a lower carbon sequestration rate with the value lower by 70.1% compared to the MF0. Based on the above results we suggested that Mv could partially replace chemical fertilizers(e.g., 40–60%) to improve or maintain the productivity and sustainability of the double-rice cropping system in South-Central China.     

Movement and survival of corn rootworm in seed mixtures of SmartStax® insect-protected corn

SmartStax^® insect-protected corn (Zea mays L.) contains genes for six Bacillus thuringiensis (Bt) proteins controlling both lepidopteran pests and the corn rootworm complex (Diabrotica spp.). The properties of SmartStax, particularly the multiple effective modes of action (i.e., each Bt protein provides a high level of control of the target pests with a low probability of cross-resistance among the proteins), have provided the opportunity to add to previously approved structured refuge options by combining the non-Bt refuge seed with SmartStax seed in a seed mix. Seed mixes ensure that a refuge is present in every Bt field, remove concerns about grower compliance with refuge requirements, and provide grower convenience. However, seed mixes could increase the likelihood that larval insects move between Bt and non-Bt plants and vice versa. Assessing the insect resistance management (IRM) value of a seed mix refuge requires an assessment of the amount of larval movement, and the consequences of that movement, for the key target pests. The studies here present such data for control of corn rootworm by SmartStax corn, which contains the rootworm-active protein Cry3Bb1 and the binary protein Cry34Ab1/Cry35Ab1. In a growth chamber experiment, SmartStax was most effective against first instars and significantly effective against second instars, but did not control third instars. In a field study of movement from a heavily infested non-Bt plant onto surrounding plants, a larger percentage of insects successfully dispersed from the infested plant when the surrounding plants were non-Bt plants than when they were SmartStax plants. A paired-plant study showed that few larvae migrated from infested SmartStax plants and survived on nearby non-Bt plants; larvae that migrated from infested non-Bt plants had low survival if the adjacent plants were SmartStax. Replicated field studies of plant-to-plant movement indicated that the non-Bt plants in a 5% or 10% seed mix consistently supported large populations of susceptible insects and represented a productive refuge, whereas the SmartStax plants had few or no survivors. The timing of emergence from seed mix plots containing 5% or 10% non-Bt plants was more similar to that of the non-Bt plots than that of the SmartStax plots. Thus, the available growth chamber and field data indicate that a seed mix of 5% or more will provide an effective refuge for corn rootworm in SmartStax corn.     

沙葱乙醇提取物对肉仔鸡生产性能的影响

试验选择120只1日龄罗斯308商品肉鸡随机分为4个处理组,每个处理3个重复,每个重复10只鸡。对照组饲喂基础日粮,试验组分为AWEⅠ组、AWEⅡ组和AWEⅢ组,分别在基础日粮中添加100、300和500 mg/kg沙葱乙醇提取物(AWE),研究其对肉仔鸡生产性能的影响。结果表明,沙葱乙醇提取物对肉仔鸡生产性能无显著影响(P0.05),但有促进日增重、提高采食量及降低料肉比等指标的趋势。     

小麦TaCYP78A5基因的克隆及生物信息学分析

细胞色素P450(Cytochromes P450)蛋白家族含有硫羟基和血红素结构域,参与多种生物合成,CYP78A家族成员对籽粒大小形成有重要的功能。利用同源克隆的方法,分别从小麦大粒品系‘P271’和小粒材料‘中国春’中扩增到位于2AS、2BS、2DS上的TaCYP78A5-2A、TaCYP78A5-2B和TaCYP78A5-2D基因,编码534、544、549个氨基酸。通过分析发现在‘P271’和‘中国春’克隆到的TaCYP78A5序列一致,说明TaCYP78A5基因在这2个粒型的小麦中是保守的。CYP78A家族在水稻和拟南芥中具有非自主性细胞表达模式,结合对CYP78A家族的生物进化关系研究发现,CYP78A家族的各个基因在单子叶植物中是保守的。‘P271’和‘中国春’籽粒大小的差异,可能是TaCYP78A5基因在‘P271’中的基因表达量高于‘中国春’,引起种子发育过程中内表皮细胞的增殖,从而促进种子表皮增大,使得‘P271’种子大小和质量增加。     

悦来航电枢纽库坝区渗漏问题的分析与评价

根据松花江悦来航电枢纽工程的勘察实例,对悦来航电枢纽库坝区渗漏问题进行了分析与评价,并且较详细地介绍了工程防治处理措施。     

栀子、龙胆、茵陈水煎剂对獭兔胆汁分泌和胆囊平滑肌运动调节研究

为了研究栀子、龙胆、茵陈水煎剂对獭兔胆汁分泌和离体胆囊平滑肌肌条运动的影响，通过对獭兔实施胆管插管术，建立胆汁引流模型，应用 BL-420E 生物机能实验系统观察记录上述中药对胆汁分泌的影响；并取獭兔胆囊平滑肌，应用 HW-400S 恒温平滑肌槽和 BL-420E 生物机能实验系统，观察记录上述中药对胆囊平滑肌运动的影响。结果显示，栀子水煎剂可显著增加胆汁的分泌（P ＜0．05）和胆囊平滑肌收缩张力（P ＜0．05）；龙胆和茵陈水煎剂可显著增加胆汁的分泌（P ＜0．05），但对胆囊平滑肌收缩张力作用不显著（P ＞0．05）。说明栀子、龙胆、茵陈水煎剂均可促进獭兔胆汁分泌，且栀子可提高胆囊平滑肌收缩张力，但龙胆、茵陈水煎剂对胆囊平滑肌收缩张力影响不显著。     

Antibiotic resistance,ESBL genes,integrons, phylogenetic groups and MLVA profiles of Escherichia coli pathotypes isolated from patients with diarrhea and farm animals in south-east of Iran

The aims of this study were to investigate the prevalence, antibiotic resistance, presence of class 1 and 2 integrons, Extended Spectrum β-Lactamases (ESBL) genes, phylogenetic group and epidemiological relationships of EPEC, ETEC and EHEC pathotypes isolated from patients with diarrhea and farm animals in south east region of Iran. A total of 671 diarrheagenic E. coli (DEC) were collected from stool samples of 395 patients with diarrhea and 276 farm cattles and goats. Presence of EPEC, ETEC and EHEC were identified using multiplex-PCR employing primers targeted the shiga toxin (stx), intimin (eae), bundle forming pili (bfp), and enterotoxins (lt and st) genes. The highest proportion of the patients (64%) were children under age 1–15 year (p ≤ 0.05). Among the isolates, atypical EPEC was detected in 26 patients and 14 animal stool samples, while typical EPEC was found in 2 cattles. ETEC isolates were detected in stools of 13 patients and 4 EHEC was identified in 3 goats and one cattle. The isolates were checked for susceptibility to 14 antibiotics. 50% (n = 13) of EPEC and 61.5% (n =8) of ETEC showed multi-drug resistance (MDR) profiles and one EPEC was found to be extensive drug resistant (XDR). In contrast, EHEC isolates were susceptible to the majority of antimicrobial agents. The MDR isolates were positive for bla_TEM and bla_CTX-M ESBL genes and carried class 1 integrons. Further study on the biofilm formation indicated that, 3 out of 4 EHEC isolates showed strong biofilm, while other pathotypes had either moderate, weak or no biofilm activity. Majority of EPEC isolates were belonged to phylogenetic group B1, all except one ETEC were classified as phylogenetic group A and two EHEC were belonged to phylogroup D, respectively. A multilocus variable tandem repeat analysis (MLVA) exhibited 22 distinct patterns. In conclusion, MLVA data showed high clonal diversity. Presence of EHEC in animal origins pose public health concern in this region.     

烟草Hsc70-2的克隆及对马铃薯Y病毒侵染烟草的促进作用

【目的】马铃薯Y病毒（Potato virus Y,PVY）是危害烟草、马铃薯、番茄、辣椒等主要农作物的重要病毒,给农业生产带来巨大损失。论文旨在克隆Hsc70-2蛋白在本氏烟（Nicotiana benthamiana）中的基因序列并分析其生物学信息,研究NbHsc70-2蛋白对PVY侵染烟草的影响,为进一步解析PVY的侵染机制提供理论依据。【方法】以本氏烟为材料,克隆NbHsc70-2蛋白的基因编码序列（coding sequence,CDS）,利用MEGA 6.0 进行多序列比对并构建系统进化树;利用实时荧光定量PCR（qRT-PCR）分析NbHsc70-2在本氏烟各组织中的表达水平;采用在线软件BaCeILo和SignalP 4.0对NbHsc70-2进行生物信息学分析;构建NbHsc70-2-RFP融合蛋白确定该蛋白的亚细胞定位;研究PVY处理对本氏烟叶片中NbHsc70-2的影响;利用激光共聚焦观察PVY-GFP处理对NbHsc70-2-RFP定位的影响;构建NbHsc70-2 VIGS沉默体系及瞬时表达载体,采用qRT-PCR比较NbHsc70-2在沉默/过表达后对PVY表达的影响。【结果】NbHsc70-2编码649个氨基酸,系统进化树分析表明,NbHsc70-2与普通烟NaHsc70-2序列相似性最高,亲缘关系最近,属于热激蛋白家族,C端具有热激蛋白家族的高度保守基序结构;qRT-PCR分析表明,NbHsc70-2在叶中表达量最高,在根和茎中的表达量较低;BaCeILo预测及激光共聚焦显微镜观察均显示NbHsc70-2定位在细胞质中,并且PVY-GFP侵染本氏烟后NbHsc70-2-RFP部分转移至细胞核,与PVY-GFP共定位在细胞质及细胞核中;将NbHsc70-2基因沉默载体pTRV::NbHsc70-2导入本氏烟中7 d后相比于对照组,沉默组出现心叶皱缩及矮化现象;接种PVY-GFP 7 d后通过手持紫外灯观察到沉默组中无明显绿色荧光出现,而对照组中PVY-GFP系统侵染使得非接种叶片呈现荧光现象,沉默组荧光点数约为对照组28%;接种PVY后,利用qRT-PCR检测沉默NbHsc70-2后1、3、5 d的PVY CP基因积累量,结果表明沉默NbHsc70-2后PVY CP基因积累量下降,其中3 d和5 d与对照相比差异显著,基因表达水平分别为对照的14%和0.004%;将NbHsc70-2过表达载体pEarleyGate100::GWC::NbHsc70-2与PVY同时导入本氏烟后,结果表明相比于对照组,过表达组48 h和72 h 其PVY CP基因积累量显著上升,基因表达水平分别为对照组的2.31、2.56倍。【结论】PVY侵染会引起NbHsc70-2表达量上升;NbHsc70-2是PVY侵染本氏烟重要的组成成分,沉默该基因显著抑制PVY的表达,过表达NbHsc70-2则显著提高PVY的表达,即NbHsc70-2的表达水平与PVY复制呈正相关,NbHsc70-2蛋白促进PVY对烟草的侵染。