Isorhamnetin protects porcine oocytes from zearalenone-induced reproductive toxicity through the PI3K/Akt signaling pathway

Background Zearalenone(ZEA)widely exists in moldy grains,which seriously destroys the fertility of females.Isorhamnetin,a natural flavonoid,has extensive of pharmacological activities.However,the beneficial effect and the underlying molecular mechanism of isorhamnetin involvement in ZEA-induced porcine oocyte damage have not been investigated.Methods Oocytes were treated with different concentrations of ZEA(3,5,8 and 10μmol/L)and isorhamnetin(5,10,20 and 30μmol/L)for 44 h at 39℃.ZEA(5μmol/L)and isorhamnetin(10μmol/L)were selected for subsequent studies.Polar body exclusion rate,apoptosis rate and apoptosis related proteins,ROS levels and SOD2 protein,mitochondrial membrane potential and distribution,endoplasmic reticulum distribution and proteins expression,and PI3K,Akt and p-Akt proteins expression of oocytes were detected.In addition,the effect of PI3K antagonist(LY294002)on oocyte nuclear maturation and apoptosis were used to determine the involvement of PI3K/Akt signaling pathway.Results Our findings showed that ZEA exposure damaged oocytes and isorhamnetin therapy restored the developmental capability of porcine oocytes.Isorhamnetin promoted polar body extrusion rate to rescue ZEA-induced meiotic arrest in porcine oocytes.Isorhamnetin alleviated ZEA-induced oxidative stress by stimulating SOD2 protein expression and inhibiting ROS production.Moreover,isorhamnetin enhanced normal mitochondrial distribution and mitochondrial membrane potential to prevent mitochondrial dysfunction induced by ZEA.Changing the expression of endoplasmic reticulum stress-related marker proteins(CHOP,GRP78)and the distribution rate of normal endoplasmic reticulum showed that isorhamnetin relieved ZEA-caused endoplasmic reticulum stress.Mechanistically,isorhamnetin decreased Bax/Bcl-2 protein expression and inhibited ZEA-induced apoptosis through PI3K/Akt signaling pathway.Conclusions Collectively,these results suggest that isorhamnetin protects oocytes from ZEA-caused damage through PI3K/Akt signaling pathway,which enhances meiotic maturation and mitochondrial function,and inhibits early apoptosis,oxidative stress and endoplasmic reticulum stress in porcine oocytes.Our study provides a new strategy for solving the reproductive toxicity induced by ZEA and treating woman infertility.

Adsorption behavior of activated carbon for the elimination of zearalenone during bleaching process of corn oil

Zearalenone is a mycotoxin produced by Fusarium species.It frequently contaminates cereals used for foods or animal feeds,especially deposited in crude corn oil.Certain amounts of zearalenone can be removed during refining processes.In this study,we studied the influence of activated carbon and six industial absorbents(zeolite,diatomite,attapulgite,perlite,montmorillonite and activated clay)on the elimination of zearalenone during bleaching process of corn oil and explored the absorption mechanism of activated carbon.Results showed that activated carbon had an excellent adsorption capacity of zearalenone compared with the other six industrial adsorbents.For activated carbon,a high removal rate of zearalenone(exceeding 83%)from heavily zearalenone-polluted corn oil was achieved and the removal rate of zearalenone was kept above 60%after five regeneration cycles.The research on the adsorption mechanism of activated carbon showed that Freundlich adsorption isotherm model and pseudo-second-order kinetic model could well described the adsorption process.The thermodynamic study demonstrated that adsorption process was spontaneous and exothermic.Fourier transform infrared spectroscopy and Raman spectroscopy further revealed that activated carbon was effectively combined with zearalenone viaπ-πinteraction.Thus,activated carbon is an efficient and suitable adsorbent to control the levels of zearalenone during bleaching process of corn oil.This study not only proposed a systematic research scheme for the mechanism study of activated carbon for the elimination of zearalenone in corn oil,but also provided the scientific basis for developing effective methods to eliminate zearalenone in refined vegetable oils.

禾谷镰孢菌产玉米赤霉烯酮毒素zearalenone的PCR检测

玉米赤霉烯酮(zearalenone,ZEN)是由镰孢菌产生的真菌毒素,该毒素具有雌性激素生物活性。PKS4基因是禾谷镰孢菌中玉米赤霉烯酮生物合成途径中的必需基因。根据PKS4基因序列,设计一对PKS4基因特异性引物,通过检测PKS4基因的存在间接检测ZEN毒素的产生。特异引物在禾谷镰孢菌菌株以及被禾谷镰孢菌侵染的小麦籽粒中都能稳定地扩增出大小为1 076 bp的特异片段,扩增片段与PKS4基因(DQ019316)相同区段序列相似性达99.63%。同时,酶联免疫吸附法(ELISA)验证了PCR的检测结果,证明了该技术的可靠性。

Aflatoxin B1, zearalenone and deoxynivalenol in feed ingredients and complete feed from different Province in China

Background： The current study was carried out to provide a reference for monitory of aflatoxin B_1（AFB_1）,zearalenone（ZEN） and deoxynivalenol（DON） contamination in feed ingredients and complete feeds were collected from different Province in China from 2013 to 2015.Methods： A total of 443 feed ingredients, including 220 corn, 24 wheat, 24 domestic distillers dried grains with soluble（DDGS）, 55 bran, 20 wheat shorts and red dog, 37 imported DDGS, 34 corn germ meal and 29 soybean meal as well as 127 complete feeds including 25 pig complete feed（powder）, 90 pig complete feed（pellet）, six duck complete feed and six cattle complete feed were randomly collected from different Province in China,respectively, by high-performance chromatography in combined with UV or fluorescence analysis.Results： The incidence rates of AFB_1, ZEN and DON contamination of feed ingredients and complete feeds were80.8, 92.3 and 93.9 %, respectively. The percentage of positive samples for DON ranged from 66.7 to 100 %.Domestic DDGS and imported DDGS presented the most serious contamination AFB_1, ZEN and DON contamination levels of feeds ranged from 61.5 to 100 %, indicated that serious contamination over the studied 3-year period.Conclusion： The current data provide clear evidence that AFB_1, ZEN and DON contamination of feed ingredients and complete feeds in different Province in China is serious and differs over past 3-year. The use of corn, domestic DDGS, imported DDGS and corn germ meal, which may be contaminated with these three mycotoxins, as animal feed may triggered a health risk for animal. Feeds are most contaminated with DON followed by ZEN and AFB_1.Mycotoxins contamination in feed ingredients and complete feeds should be monitored routinely in China.

Alleviation of mycotoxin biodegradation agent on zearalenone and deoxynivalenol toxicosis in immature gilts

Background: The current study was carried out to evaluate the effects of mycotoxin biodegradation agent(MBA, composed of Bacillus subtilis ANSB01 G and Devosia sp. ANSB714) on relieving zearalenone(ZEA) and deoxynivalenol(DON) toxicosis in immature gilts.Methods: A total of forty pre-pubertal female gilts(61.42 ± 1.18 kg) were randomly allocated to four diet treatments: CO(positive control); MO(negative control, ZEA 596.86 μg/kg feed and DON 796 μg/kg feed);COA(CO + 2 g MBA/kg feed); MOA(MO + 2 g MBA/kg feed). Each treatment contained 10 replicates with 1 gilt per replicate. Gilts were housed in an environmentally controlled room with the partially slatted floor.Results: During the entire experimental period of 28 d, average daily gain(ADG) and average daily feed intake(ADFI)of gilts in MO group was significantly reduced compared with those in CO group. The vulva size of gilts was significantly higher in MO group than CO group. In addition, significant increases in the plasma levels of Ig A,Ig G, IL-8, IL-10 and PRL were determined in MO group compared with that in CO group. ZEA and DON in the diet upregulated apoptotic caspase-3 in ovaries and uteri, along with down-regulated the anti-apoptotic protein Bcl-2 in ovaries. The supplementation of MBA into diets co-contaminated with ZEA and DON significantly increased ADG, decreased the vulva sizes, reduced the levels of Ig G, IL-8 and PRL in plasma, and regulated apoptosis in ovaries and uteri of gilts.Conclusions: The present results indicated that feeding diet contaminated with ZEA and DON simultaneously(596.86 μg/kg + 796 μg/kg) had detrimental effects on growth performance, plasma immune function and reproductive status of gilts. And MBA could reduce the negative impacts of these two toxins, believed as a promising feed additive for mitigating toxicosis of ZEA and DON at low levels in gilts.

Time-resolved fluoroimmunoassay of zearalenone in cereals with a europium chelate as label

A competitive indirect time-resolved fluoroimmunoassay(TRFIA) was developed for detection of zearalenone(ZEN) in cereals,in which ZEN conjugated to bovine serum albumin(BSA) is used as solid-phase antigen.A competitive indirect TRFIA was conducted by simultaneously incubating ZEN in standard or extracted samples with anti-ZEN monoclonal antibody over ZEN-BSA coated plates,and then determining the bound ZEN monoclonal antibody with goat anti-mouse europium conjugate.Samples were extracted with methanol/water...

Modified halloysite nanotubes reduce the toxic effects of zearalenone in gestating sows on growth and muscle development of their offsprings

Background： Zearalenone （ZEN） is an estrogenic mycotoxin that is primarily produced by Fusarium fungi and has been proven to affect the reproductive capacity of many species to varying degrees. The present experiment was designed to study the maternal persistent effects of zearalenone toxicity in gestating sows on growth and muscle development of their offsprings, and the alleviation of zearalenone toxicity by modified halloysite nanotubes （MHNTs）. Methods： Eighteen sows were fed with one of three dietary treatments that included the following： （1） a control diet, （2） a contaminated grain diet （with 50 % moldy corn, 2.77 mg/kg ZEN）, and （3） a contaminated grain diet （with 50 % moldy corn, 2.76 mg/kg ZEN） ＋ 1% MHNTs. Each sow was exclusively fed its experimental diets from 35 to 70 d of gestation at a total of 2 kg daily. Muscle samples were collected from six piglets per treatment at birth, weaning and finishing. Results： The results showed that feeding the sows with the ZEN-contaminated diets from 35 to 70 d of gestation decreased the ADG, ADFI and G：F of their offsprings （P 〈 0.05）. The muscle fiber numbers in the newborn, weaning and growing-finishing pigs and the muscle fiber diameters at birth and weaning were also decreased by maternal ZEN exposure （P 〈 0.05）. The expressions of IGF-I, IGF-II, Myf-5 and Mstn at birth and IGF-II, Pax7, Myf-5 and MyoD1 at weaning were altered by feeding gestating sows with ZEN-contaminated diets （P 〈 0.05）. The MHNTs reduced most of the ZEN-induced toxic effects： the ADG and ADFI on growth performance, the muscle fiber numbers at weaning and finishing and the muscle fiber diameters at weaning （P 〈 0.05）. The expression levels of IGF-II and Mstn in newborn piglets and IGF-II and Myf-5 in weaning piglets were also prevented by adding 1% MHNTs （P 〈 0.05）. Conclusions： The present study demonstrated that the offsprings of sows fed with ZEN-contaminated diets from 35 to 70 day of gestation exhibited weakening on growth performance, physiological changes in their muscle fibers and alterations of mRNA expression in their muscle tissues, and also indicated that MHNTs prevented most of the ZEN- induced weakening in the muscle tissues.

Occurrence of Aflatoxin B_(1),deoxynivalenol and zearalenone in feeds in China during 2018-2020

Background:The current study was conducted to investigate the individual and combined occurrence of aflatoxin B_(1)(AFB_(1)),deoxynivalenol(DON)and zearalenone(ZEN)in feeds from various Provinces of China during 2018 to 2020.A total of 3,507 feed samples,including 2,090 feed ingredients and 1,417 complete feed samples,were collected from different areas of China for mycotoxins analysis.Results:The individual contamination of AFB_(1),DON and ZEN were present in more than 81.9%,96.4% and 96.9% of feed samples,respectively,with average concentration ranges of AFB_(1) between 1.2-27.4μg/kg,DON between 458.0-1,925.4μg/kg and ZEN between 48.1-326.8μg/kg.Notably,0.9%,0.5% and 0.1% of feed ingredients,and 1.2-12.8%,0.9-2.9% and 0-8.9% of complete feeds for pigs,poultry and ruminants with AFB_(1),ZEN and DON that exceeded China’s safety standards,respectively.Moreover,more than 81.5%of feed ingredients and 95.7% of complete feeds were co-contaminated with various combinations of these mycotoxins.Conclusion:This study indicates that the feeds in China were universally contaminated with AFB_(1),DON and ZEN during the past 3 years.These findings highlight the significance of monitoring mycotoxin contaminant levels in the domestic animal feed,and the importance of carrying out feed administration and remediation strategies for mycotoxin control.

Recent advances in detoxification strategies for zearalenone contamination in food and feed

Zearalenone(ZEN)is a widely distributed mycotoxin that frequently contaminates crops and animal feed.ZEN can cause serious health problems in livestock and humans alike,leading to great economic losses in the food industry and livestock farming.Therefore,approaches for efficient ZEN decontamination in food and feed are urgently needed.Traditional physical and chemical methods may decrease the nutritional quality of food and palatability of feed,or leading to residues and safety concerns.By contrast,biological methods for the removal or degradation of ZEN overcome these problems,especially for biological degradation by microorganisms and specific enzymes extracted from strains that can convert ZEN to less toxic or even completely harmless products.In this review,we comprehensively describe methods for ZEN degradation,focusing especially on biological strategies.Finally,emerging strategies and advice on remaining challenges in biodegradation research are also briefly discussed.

Investigation of the Biochemical and Histological Changes Induced by Zearalenone Mycotoxin on Liver in Male Mice and the Protective Role of Crude Venom Extracted from Jellyfish <i>Cassiopea Andromeda</i>

Zearalenone (ZEN) is a non steroidal estrogenic mycotoxin produced by Fusarium species of fungi which contaminate human foods and animal feeds worldwide. In this study hepatotoxicity of ZEN was evaluated in mice by oral adminis-tration of single and repeated doses of ZEN mycotoxin (2.7 mg/kg b.w.). The protective effect of crude venom extracted from jellyfish Cassiopea andromeda was also assessed. Mice were divided into four groups (N = 10). G1: receiving the toxin once and sacrificed 48 h later, G2: toxin administered twice for one week, G3: toxin administered twice a week for two weeks, G4: pretreated orally by a single dose of crude venom (1.78 mg/20g) 24 hours prior to administration of ZEN twice a week for two weeks. Each treated group had its corresponding control which received 1% DMSO sa-line.ZEN treatment significantly increased alanine aminotransferase (ALT), aspartateaminotrnsferase (AST) and alka-line phosphatase (ALP) activities after 48 hours and two weeks, while ALT was also significantly increased after one week. Tumor necrosis factoralpha (TNF-α) level was undetected in treated and control groups except the group treated with ZEN for one week. Alphafetoprotein (AFP) level was increased significantly only after two weeks. The activity of antioxidants was significantly increased in all groups. ZEN was also found to modify the serum proteins especially gamma-globulin which showed a significant decrease after 48 h and two weeks. Improvement in liver func-tion occurred in the group pretreated with the crude venom, and AFP and antioxidants returned to normal level, while TNF-α level was also undetected. Gamma globulin was significantly increased. The recovery observed in the group which was pretreated with crude venom may related to bradykinin content of this venom which exhibits a hepatoprotective effect. Histological changes in mouse liver coincided with biochemical changes. In conclusion, this study revealed that ZEN induced liver function and structural changes promising an approach for using a crude venom of jellyfish to enhance liver function.

Review on microbial degradation of zearalenone and aflatoxins

The widespread contamination by mycotoxins,especially zearalenone and aflatoxins,in crops and their by-products has caused severe undesirable effects on human health and commercial trade.Researchers had screened out microorganisms from various media for degrading zearalenone and aflatoxins,and the results of a lot of studies showed that enzymes derived from microbial strains play a key role in degrading mycotoxins.Genetic engineering technology had been applied to improve the heterologously expressed degrading enzymes in several mature microbial hosts such as Escherichia coli and Pichia pastoris.The separated and purified recombinant enzyme had high activity in degrading mycotoxins in vitro.This review summarized the types of mycotoxins-degrading microorganisms and enzymes,and the progress on synthesis of heterologously expressed degrading enzymes by genetic engineering technology as well as related researches on improving the effect of degrading enzymes.We also prospected the future development in the study of using recombinant enzymes formed by genetic engineering technology to realize the simultaneous degradation of multiple mycotoxins in crops.

IDENTIFICATION OF ALTERNARIOL, ALTERNARIOL MONOMETHYL ETHER AND ZEARALENONE BY PARTICLE BEAM LC/MS

A method for simultaneously analyzing altemariol(AOH), altemariol monomethyl ether (AME) and zearalenone(ZEA) by particle beam LC/MS was established, LC separation was accompiished with a solvent system of methanol and water (80:20 v/v). The followed particle beam LC/MS analysis gave searchable spectra of AOH. AME and ZEA. Application of this technique to analysis of an alternaria culture confirmed the presence of AOH and AME.

姜黄素介导光动力抑制禾谷镰刀菌产ZEN毒素的研究

玉米赤霉烯酮(Zearalenone,ZEN)是广泛污染玉米、小麦、大米等谷物的真菌毒素之一,主要由镰刀菌属产生。该研究选择姜黄素为光敏剂构建光动力技术(Photodynamic Inactivation,PDI),探究姜黄素浓度、光照能量和溶液pH对PDI抑制禾谷镰刀菌生长及其在玉米中产ZEN的影响。同时,从ZEN毒素含量变化和禾谷镰刀菌失活两个角度分析PDI抑制禾谷镰刀菌产ZEN的机制。结果表明,在50.00μmol/L姜黄素和43.0 J/cm^(2)光照能量作用下,ZEN含量从空白对照玉米中的1.15 mg/kg降低至光动力处理组中的0.58 mg/kg,降幅达49.56%,且继续增加姜黄素浓度能够完全抑制ZEN的生成。深入分析表明,PDI对禾谷镰刀菌的灭活作用是其抑制玉米中ZEN产生的主要原因,尤其当姜黄素浓度≥70.00μmol/L,光照能量为43.0 J/cm^(2)时,禾谷镰刀菌被完全灭活。因此,该研究证实光动力技术能够有效地杀灭禾谷镰刀菌,抑制ZEN的产生,为保障农产品质量安全提供了一种新方法。

A Study on the“Social Behavior”of Zen Poetry Culture:A Case Study of the Sui and Tang Dynasties

During the Sui and Tang dynasties,Buddhism and Zen poetry underwent great development.Zen poetry is diverse in form,strong in personality,and high in achievement.Among the types of Zen poetry are poems,lyrics,and prose,some of which have social attributes closely related to the development of Chinese Buddhism.An analysis reveals that the viewpoints and information conveyed in these poems,lyrics,and prose are positively correlated with the attitudes of the emperors towards Buddhism.When the attitude of the emperor towards Buddhism reached its peak of positivity during the Tang dynasty,there was a change towards a negative correlation with the viewpoints expressed in poetry.

Electrochemical Reduction of Zearalenone

Electrochemical reduction of zearalenone 1 and its derivatives was described. The ratio of a-and b-zearalanol 2 obtained by electrochemical reduction reached 8:1, much higher tan that obtained from catalytic hydrogenation. In addition to of the normal reduction products a rearranged product 5 was isolated. The reduction was supposed to be a electrocatalytical reaction.

基于金纳米花的双信号适配体传感器检测红酒中真菌毒素

构建基于金纳米花(gold nanoflower,AuFL)和适配体的双信号荧光适配体传感器用于同时检测赭曲霉毒素A(ochratoxin A,OTA)和玉米赤霉烯酮(zearalenone,ZEN)。以AuFL为载体,将含OTA和ZEN适配体的单链DNA共价交联在AuFL表面,进一步将其与两种荧光染料(FAM和Cy3)标记的互补DNA杂交,制备DNA功能化纳米探针。荧光染料和AuFL之间发生荧光共振能量转移(fluorescence resonance energy transfer,FRET)导致纳米探针自身无荧光。加入OTA和ZEN后,FAM和Cy3从探针表面脱落会扰乱FRET过程,导致两者荧光的恢复。优化条件下,该双信号荧光适配体传感器对OTA检测的线性范围为0.05~500 ng/mL,检出限为0.017 ng/mL。对ZEN检测的线性范围为0.1~500ng/mL,检出限为0.033ng/mL。该双信号荧光适配体传感器具有良好的选择性和重现性,并能够成功应用于红酒实际样中OTA和ZEN的检测。

白藜芦醇对ZEN诱导的猪小肠上皮细胞损伤的作用效果研究

试验旨在探究白藜芦醇对玉米赤霉烯酮(ZEN)诱导的猪小肠上皮细胞(IPEC-J2细胞)损伤的影响。试验分组为:C组(对照组)、Z组(ZEN浓度25µg/mL)、ZR1组(25µg/mL ZEN+10µmol/L白藜芦醇)、ZR2组(25µg/mL ZEN+20µmol/L白藜芦醇)、ZR3组(25µg/mL ZEN+30µmol/L白藜芦醇);测定细胞活力、细胞乳酸脱氢酶(LDH)活性、抗氧化酶水平以及Caspase 3、Caspase9、Bcl-2和bax等凋亡相关基因mRNA表达水平。试验结果显示:Z组细胞活力、SOD活性T-AOC水平和Bcl-2基因mRNA表达水平极显著低于C组(P<0.01),ZR2组和ZR3组细胞活力、SOD活性、T-AOC水平和Bcl-2基因mRNA表达水平极显著或显著高于Z组和ZR1组(P<0.01或P<0.05);Z组细胞LDH活性、MDA水平、Capase-3、Capase-9和Bax基因mRNA表达水平极显著高于C组(P<0.01),ZR2组和ZR3组细胞LDH活性、MDA水平、Capase-3、Capase-9和Bax基因mRNA表达水平极显著或显著低于Z组和ZR1组(P<0.01或P<0.05),ZR1组细胞LDH活性和Bax基因mRNA表达水平极显著低于Z组(P<0.01)。结果表明,白藜芦醇能够缓解ZEN诱导的IPEC-J2细胞的细胞活力、LDH活性、抗氧化功能和凋亡相关基因mRNA表达水平的改变。

高效液相荧光色谱法同时测定六神曲中黄曲霉毒素、玉米赤霉烯酮和赭曲霉毒素A

目的建立一种用免疫亲和柱净化-柱后光化学衍生-高效液相色谱同时检测六神曲中黄曲霉毒素、玉米赤霉烯酮和赭曲霉毒素A的方法。方法样品采用60%乙腈超声提取,免疫亲和柱净化,采用XBridge^(®)Phenyl苯基色谱柱(4.6 mm×250 mm,5μm),以乙腈和0.1%磷酸溶液为流动相,梯度洗脱,光化学衍生仪衍生,通过切换荧光波长检测。结果黄曲霉毒素、玉米赤霉烯酮和赭曲霉毒素A标准曲线的线性范围分别为0.006~0.108、0.500~2.500和0.202~1.009 ng,6种毒素的线性关系在0.9994以上,检出限分别为1.2、100.0和40.3 ng/ml,平均加标回收率为73.2%~92.3%,相对偏差为2.1%~5.4%。结论该方法具有专属性强、操作方便等特点,能够有效用于六神曲中3种毒素的同时检测和安全质量控制。

中国美学的两次突破理论及其方法论反思——关于《禅宗美学》的对话

张节末教授的代表作之一《禅宗美学》,综合使用突破法与会通法,融汇康德美学与现象学还原法,采取哲学美学立场,创新性地研究了中国美学史上从庄子、玄学到禅宗的美学突破。分析最基础的禅之感性经验,提出禅宗主空幻的自然观与看空的人格观相结合,催生心化的境界—意境的审美经验体系。多年后反思《禅宗美学》的方法论,提出美学研究必需保持理论思维的哲学高度,同时要下沉到文物和艺术,向美学史贡献历史感与现实感。

夏目漱石汉诗中的禅宗审美意蕴呈现及其原因研究

夏目漱石在创作汉诗时巧妙融入人生品味理念,自然天成禅宗美学意蕴,创造出日本本土化汉诗文的样态,借此实现中国传统文化在日本的语境转换。本稿在分析了夏目漱石汉诗文中的禅宗理念基础上,然后以具体示例重点阐释了其汉诗文中所蕴含的“物哀、幽玄”之禅宗意境、“动静”合璧之禅画意境、“道”之禅宗神韵等审美内涵。最后,从夏目漱石深厚的汉文学修养功底、艰辛坎坷的个人经历以及日本传统文化审美意识的影响等方面,详细分析了夏目漱石汉诗文中所蕴含的禅宗审美思想之来源。