Bacterial infections of avian embryos can lead to an increase in embryo mortality,and the proliferation of antimicrobial-resistant bacteria aggravates the situation.A low hatching rate also poses a challenge to the po...Bacterial infections of avian embryos can lead to an increase in embryo mortality,and the proliferation of antimicrobial-resistant bacteria aggravates the situation.A low hatching rate also poses a challenge to the population of artificially bred Crested Ibises(Nipponia nippon).This study aims to determine the potential association between bacterial infection and the death of Crested Ibis embryos,and whether there is convergence between antimicrobial resistance and virulence in strain.In this study,13 Escherichia coli and 12 Proteus mirabilis isolates were recovered from dead Crested Ibis embryos.The pathogenicity examination confirmed the pathogenicity of all isolates,and multiple virulence genes detected by PCR-sequencing demonstrated the presence of irp2 and iuc D(100%),fim C and iss(92.31%)in E.coli,and uca A(58.33%)in P.mirabilis.Antimicrobial susceptibility test demonstrated that isolates were mainly resistant to amoxicillin(E.coli:76.92%,P.mirabilis:91.67%),cefazolin(E.coli:76.92%,P.mirabilis:91.67%),oxytetracycline(E.coli:92.31%,P.mirabilis:75.00%)and sulfamethoxazole-trimethoprim(E.coli:53.85%,P.mirabilis:33.33%),and more than 30%of isolates showed multidrug-resistance(MDR).Further analyses detected extended-spectrumβ-lactamase(ESBL)genes,of which blaTEM-1(E.coli:100%,P.mirabilis:100%)had the highest frequency,followed by the blaCTX-M-55(E.coli:92.31%,P.mirabilis:50%),blaCTX-M-14(E.coli:76.92%,P.mirabilis:33.33%),blaCTX-M-65(E.coli:15.38%,P.mirabilis:16.67%),and all isolates were negative for blaSHV and blaOXA.Pearson's correlation analysis showed a positive correlation between the presence ofβ-lactam resistance and ESBL genes,while mainly negative correlations were observed between the presence of ESBL genes and virulence genes.Furthermore,the conjugation experiment and PFGE revealed that the isolates were primarily polyclonal,and there was horizontal transfer of resistance or virulence genes by plasmids.Based on the results,E.coli and P.mirabilis were responsible for embryonic mortality of the ibises in this study.The co-presence and co-transfer of ESBL genes and virulence genes can pose a potential threat to the health of the Crested Ibis,and measures such as prudent use of antimicrobials,and constant surveillance of resistance and pathogenicity,must be implemented at the Crested Ibis breeding base.展开更多
Litchi has great economic significance as a global fruit crop.However,the advancement of litchi functional genomics has encountered substantial obstacles due to its recalcitrance to stable transformation.Here,we prese...Litchi has great economic significance as a global fruit crop.However,the advancement of litchi functional genomics has encountered substantial obstacles due to its recalcitrance to stable transformation.Here,we present an efficacious Agrobacterium tumefaciens-mediated transformation system in somatic embryos of‘Heiye'litchi.This system was developed through the optimization of key variables encompassing explant selection,A.tumefaciens strain delineation,bacterium concentration,infection duration,and infection methodology.The subsequent validation of the transformation technique in litchi was realized through the ectopic expression of LcMYB1,resulting in the generation of transgenic calli.However,the differentiation of transgenic calli into somatic embryos encountered substantial challenges.To delineate the intricate molecular underpinnings of LcMYB1's inhibitory role in somatic embryo induction,a comprehensive transcriptome analysis was conducted that encompassed embryogenic calli(C),globular embryos(G),and transgenic calli(TC).A total of 1,166 common differentially expressed genes(DEGs)were identified between C-vs.-G and C-vs.-TC.Gene Ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis revealed that these common DEGs were mostly related to plant hormone signal transduction pathways.Furthermore,RT-qPCR corroborated the pronounced down-regulation of numerous genes that are associated with somatic embryo induction within the transgenic calli.The development of this transformation system provides valuable support for functional genomics research in litchi.展开更多
Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.There...Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos.展开更多
As an increasingly used alternative to perfluorooctanoic acid(PFOA),hexafluoropropylene oxide trimer acid(HFPO-TA)has been widely detected in global water environments.However,little is known regarding its toxic effec...As an increasingly used alternative to perfluorooctanoic acid(PFOA),hexafluoropropylene oxide trimer acid(HFPO-TA)has been widely detected in global water environments.However,little is known regarding its toxic effects on cardiovascular development.Here,zebrafish embryos were treated with egg water containing 0,60,120,or 240 mg/L HFPO-TA.Results showed that HFPO-TA treatment led to a significant reduction in both larval survival percentage and heart rate.Furthermore,HFPO-TA exposure caused severe pericardial edema and elongation of the sinus venous to bulbus arteriosus distance(SV-BA)in Tg(myl7:GFP)transgenic larvae,disrupting the expression of genes involved in heart development and thus causing abnormal heart looping.Obvious sprouting angiogenesis was observed in the 120 and 240 mg/L exposed Tg(fli:GFP)transgenic larvae.HFPO-TA treatment also impacted the mRNA levels of genes involved in the vascular endothelial growth factor(VEGF)pathway and embryonic vascular development.HFPO-TA exposure significantly decreased erythrocyte number in Tg(gata1:DsRed)transgenic embryos and influenced gene expression associated with the heme metabolism pathway.HFPO-TA also induced oxidative stress and altered the transcriptional levels of genes related to cell cycle and apoptosis,inhibiting cell proliferation while promoting apoptosis.Therefore,HFPO-TA exposure may induce abnormal development of the cardiovascular and hematopoietic systems in zebrafish embryos,suggesting it may not be a suitable or safe alternative for PFOA.展开更多
The thermal-elastic martensitic transformation from high-temperature Ni_(2)In-type hexagonal structure to low-temperature TiNiSi-type orthorhombic structure has been widely studied in MnMX(M=Ni or Co,and X=Ge or Si)al...The thermal-elastic martensitic transformation from high-temperature Ni_(2)In-type hexagonal structure to low-temperature TiNiSi-type orthorhombic structure has been widely studied in MnMX(M=Ni or Co,and X=Ge or Si)alloys.However,the answer to how the orthorhombic martensite nucleates and grows within the hexagonal parent is still unclear.In this work,the hexagonal-orthorhombic martensitic transformation in a Co and Ge co-substituted MnNiSi is investigated.One can find some orthorhombic laths embedded in the hexagonal parent at a temperature above the martensitic transformation start temperature(M_(s)).With the the sample cooing to M_(s),the laths turn broader,indicating that the martensitic transformation starts from these pre-existing orthorhombic laths.Microstructure observation suggests that these pre-existing orthorhombic laths do not originate from the hexagonal-orthorhombic martensitic transformation because of the difference between atomic occupations of doping elements in the hexagonal parent and those in the preexisting orthorhombic laths.The phenomenological crystallographic theory and experimental investigations prove that the pre-existing orthorhombic lath and generated orthorhombic martensite have the same crystallography relationship to the hexagonal parent.Therefore,the orthorhombic martensite can take these pre-existing laths as embryos and grow up.This work implies that the martensitic transformation in MnNiSi_(1-x)(CoNiGe)_(x) alloy is initiated by orthorhombic embryos.展开更多
[Objective]This study was to screen out suitable genotypes and basic medium for the culture of maize mature embryos.[Method]Using mature embryos of nine maize genotypes as explants,the effects of genotypes and basic m...[Objective]This study was to screen out suitable genotypes and basic medium for the culture of maize mature embryos.[Method]Using mature embryos of nine maize genotypes as explants,the effects of genotypes and basic medium on callus induction and subculture were investigated.[Result]The genotypes performed better in callus induction and subculture were found in turn 853-35,853-209,Dan 34 and 81162.MS medium is better than N6 medium in the callus induction from maize embryos,while N6 medium is more suitable for callus subculture.[Conclusion]Our study further improved the tissue culture system in maize with mature embryos as explants.展开更多
In this study we studied the factors influencing the callus induction from mature embryos of maize inbred lines Qi 319, Zhen 58, Chang 7 -2, Lx 9801 and 81162, such as genotype, combination of plant growth regulators,...In this study we studied the factors influencing the callus induction from mature embryos of maize inbred lines Qi 319, Zhen 58, Chang 7 -2, Lx 9801 and 81162, such as genotype, combination of plant growth regulators, and low-temperature pretreatment. The results showed that the induction rate of Qi 319 was the highest among the four genotypes tested; combination of 4.0 mg/L 2,4-D ＋ 0.5 mg/L 6-BA was suitable for inducing callus from mature embryos; three days of 4℃ pretreatment can promote the callus induction significantly. The indices optimized in the present study are helpful for establishing genetic transformation system in maize without considering seasonal variation.展开更多
Fluctuations of levels of several endogenous plant hormones in isolated rice ( Oryza sativa ssp. japonica) embryos during early and mid-embryogenesis and early stages of germination were studied by enzyme-linked immun...Fluctuations of levels of several endogenous plant hormones in isolated rice ( Oryza sativa ssp. japonica) embryos during early and mid-embryogenesis and early stages of germination were studied by enzyme-linked immunosorbent assay (ELISA). Embryos were collected at different days after pollination (DAP) and different days after imbibition (DAI) of mature seeds. The contents of gibberellin(1) (GA(1)), abscisic acid (ABA), isopentenyladenine and isopentenyladenosine ( iPAs), zeatin and zeatin riboside ( ZRs) were immunochemically assayed. The GA(1) level was the highest among all hormones tested. The variations of GA(1) levels were opposite with the ABA levels, with some exceptions. During early and mid-embryogenesis, the levels of GA(1) and ABA were the highest at 4 DAP. From 8 to 18 DAP, GA(1) level declined, whereas the ABA level increased. During germination, GA(1) level increased at 2 DAI whereas simultaneously the ABA content decreased. The highest ratio of GA(1)/ABA was observed at 2 DAI The levels of iPAs and ZRs were maxima in the embryos at 4 DAP, decreased to a very low level and maintained constant thereafter. Our results provide further evidence that GA(1) plays an important role in the early stages of embryo development and germination. The balance between GA(1) and ABA, rather than their absolute contents, controls these processes throughout the development, whereas iPAs and ZRs may play important roles in early embryogenesis. The use of isolated embryos as starting material avoids the usual interferences with other tissues such as the endosperm. In addition, this is the first report dealing with the hormonal balance of early-embryos in rice.展开更多
An in situ hybridization technique for localization of calmodulin(CaM) mRNA in isolated entire embryo sacs and proembryos in Nicotiana tabacum L.cv.W38 has been developed. This technique can be applied to smal...An in situ hybridization technique for localization of calmodulin(CaM) mRNA in isolated entire embryo sacs and proembryos in Nicotiana tabacum L.cv.W38 has been developed. This technique can be applied to small amounts of materials in which a whole view of CaM mRNA distribution can be obtained. The authors revealed that CaM mRNA expression changes dramatically before and after fertilization. Especially interesting is that a prominent CaM mRNA band appears between the egg apparatus and polar nuclei temporarily during the period of pollination and fertilization. The band disappears just prior to fertilization and expands to a fan_shaped region that occupies the micropylar portion of the embryo sac. After fertilization, CaM mRNA accumulates in the elongated zygotes with higher concentration in their chalazal portion than in the micropylar portion. Such an asymmetrical pattern continues to manifest in the early proembryos. It is supposed that CaM mRNA may be involved in the early events and signaling steps associated with double fertilization and zygote polarization in higher plants.展开更多
In ecological zone of Chengdu, Sichuan, microspore culture was carried out in Brassica napus L. to study the influencing factors on microspore culture. The results showed that the temperature on microspore formation s...In ecological zone of Chengdu, Sichuan, microspore culture was carried out in Brassica napus L. to study the influencing factors on microspore culture. The results showed that the temperature on microspore formation stage, day and night temperature, disinfection solution of buds, cultivation concentration on microspore and strain-age were both important influencing factors on microspore culture. At a temperature below 5 ℃ or above 20 ℃, the material had a much lower embryo producing rate or even could not produce any embryo, but at the optimum temperature of 10 -15 ℃ the embryo yield was up to 300 pieces per bud; the best cultivation effect appeared when 0. 1% HgCl2 was used for disin- fection; the best density of microspore was 3 -4 buds per dish; In 2009, while strain-age was from 125 d to 150 d, the microspore embryo yield increased as strain-age increased. When stain-age was 150 days, the microspore embryo yield was up to the highest, but the yield declined after 150 days.展开更多
Polystyrene nanoparticles pose significant toxicological risks to aquatic ecosystems,yet their impact on zebrafish(Danio rerio)embryonic development,particularly erythropoiesis,remains underexplored.This study used si...Polystyrene nanoparticles pose significant toxicological risks to aquatic ecosystems,yet their impact on zebrafish(Danio rerio)embryonic development,particularly erythropoiesis,remains underexplored.This study used single-cell RNA sequencing to comprehensively evaluate the effects of polystyrene nanoparticle exposure on erythropoiesis in zebrafish embryos.In vivo validation experiments corroborated the transcriptomic findings,revealing that polystyrene nanoparticle exposure disrupted erythrocyte differentiation,as evidenced by the decrease in mature erythrocytes and concomitant increase in immature erythrocytes.Additionally,impaired heme synthesis further contributed to the diminished erythrocyte population.These findings underscore the toxic effects of polystyrene nanoparticles on hematopoietic processes,highlighting their potential to compromise organismal health in aquatic environments.展开更多
Background Increased backfat thickness of sows in early gestation is negative to reproductive performance.Endometrial receptivity is an important determinant of reproductive success,but it is unclear whether the effec...Background Increased backfat thickness of sows in early gestation is negative to reproductive performance.Endometrial receptivity is an important determinant of reproductive success,but it is unclear whether the effect of sow backfat thickness on litter size is associated with endometrial receptivity and whether melatonin treatment may have benefits.The present study seeks to answer these questions through in vitro and in vivo investigations.Results Excessive lipid deposition and lower melatonin levels in the uterus are detrimental to endometrial receptivity and embryo implantation in high backfat thickness sows.In cells treated with melatonin,the MT2/PI3K/LIF axis played a role in reducing lipid accumulation in porcine endometrial epithelium cells and improved endometrial receptivity.Furthermore,we found a reduction of lipids in the uterus after eight weeks of intraperitoneal administration of melatonin to HFD mice.Notably,melatonin treatment caused a significant reduction in the deposition of endometrial collagen,an increase in the number of glands,and repair of the pinopode structure,ultimately improving endometrial receptivity,promoting embryo implantation,and increasing the number of litter size of mice.Conclusions Collectively,the finding reveals the harmful effects of high backfat thickness sows on embryo implantation and highlight the role of melatonin and the MT2/PI3K/LIF axis in improving endometrial receptivity by enhancing metabolism and reducing the levels of uterine lipids in obese animals.展开更多
[Objective]The aim was to study the effective factors on culturing Prunus salicina cv.Zaoshi embryos in vitro.[Method]Different age,culture medium and GA3 which affected culturing of prumssalicina embryos were discuss...[Objective]The aim was to study the effective factors on culturing Prunus salicina cv.Zaoshi embryos in vitro.[Method]Different age,culture medium and GA3 which affected culturing of prumssalicina embryos were discussed.[Result]The result showed that on the part of 26-41 d,after the blooming period,the embryos remained tiny and retained endosperms and showed no signs of change after having cultured for three generations.On the part of 48 d,after the blooming period,the endosperms had disappeared,the embryos kept growing until they filled the seed cavity;when they were planted on the MS culture medium,their survival rate reached 77%,in its first generation,the response of embryos was discernible.On the part of 65 d,after the blooming period,4.5% of their embryos grew into shoots on the MS culture medium;with the age of embryos growing,the survival rate of shoots increased until it reached 26% when the fruits went into ripeness;the embryos produced calli in their first generation of culturing.On the part of 65-83 d,after the blooming period,the embryos produced calli through more than 2-3 generations.On the part of 88 d,after the blooming period,the survival rate of shoots on the WPM culturing base doubled compared with that on the MS culturing base;on the same culture medium,the embryos were inhibited from growing into shoots when BA,KT or 2,4-D was added on to the culture medium.The survival rate of shoots was increased remarkably when the seeds were treated in 1 000 mg/L GA3.[Conclusion] This study provided experimental basis for the establishment of Prunus salicina cv.Zaoshi,embryos rescue techniques and cross breading.展开更多
[Objective] The aim of this study was to manually isolate the microstructures such as ovule and embryo of Capsella bursa-pastoris (L.) Medic. by the non-enzymatic method. [Method] With the gum block and dissecting mic...[Objective] The aim of this study was to manually isolate the microstructures such as ovule and embryo of Capsella bursa-pastoris (L.) Medic. by the non-enzymatic method. [Method] With the gum block and dissecting microscope, its floral characteristics, ovule and embryo were manually isolated and observed by the non-enzymatic method. [Result] C. bursa-pastoris had four calyces, four petals, tetradynamous stamen and silicle, while its embryogenesis contained globular embryo, heart-shaped embryo and torpedo-shaped embryo. [Conclusion] The microstructures such as ovule and embryo of plants become easier to be isolated manually with the viscosity of gum block, which has small damage on embryo or disadvantages of keeping original shapes, and also can used in multi fields studies. This test can also be accomplished in labs with poor experiment facilities.展开更多
[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the eff...[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the effect of different 2,4-D concentrations on the induction of callus,the effect of different 6-BA concentrations on the differentiation of test-tube plantlet,as well as the effect of different IBA concentrations on the rooting of test-tube plantlet. [Result] 2,4-D showed obvious effect on the induction of inducement rate of maize,and the optimum induction medium was：N6 ＋ 2 mg/L of 2,4-D ＋ 500 mg/L of CH ＋ 500 mg/L of Pro ＋10 mg/L of AgNO3; the optimum differentiation medium was：N6 ＋ 0.5 mg/L of BA ＋ 500 mg/L of Pro; the optimum for the rooting of test-tube plantlet was 1/2 MS ＋ 0.5 mg/L of IBA. [Conclusion] The study had provided basis for the genetic transformation of maize.展开更多
The rapidly evolving environment of assisted reproductive technology(ART)requires consideration of how new innovations are reshaping clinical practice as much as research.In particular,there are three key areas that,w...The rapidly evolving environment of assisted reproductive technology(ART)requires consideration of how new innovations are reshaping clinical practice as much as research.In particular,there are three key areas that,while full of promise,also present significant challenges:the use of artificial intelligence(AI)in embryo selection,the impact of personalized medicine on ART success rates,and the ethical considerations of genetic screening of embryos[1].This letter is meant to provoke further discussion and highlight the need for harmonized global guidelines as these advances continue to reshape the reproductive medicine environment.展开更多
Background MicroRNAs(miRNAs)are presented in the uterine lumen of many mammals,and in vitro experiments have determined that several miRNAs are important for the regulation of endometrial and trophoblast functions.Our...Background MicroRNAs(miRNAs)are presented in the uterine lumen of many mammals,and in vitro experiments have determined that several miRNAs are important for the regulation of endometrial and trophoblast functions.Our aim was to identify and contrast the miRNAs present in extracellular vesicles(EVs)in the uterine lumen fluid(ULF)at the onset of attachment in cattle pregnancies(gestation d 18)initiated by artificial insemination(AI)or by the trans-fer of an in vitro-produced blastocyst(IVP-ET).A third group had no conceptus after the transfer of an IVP embryo.Results The abundance of 263 annotated miRNAs was quantified in the EVs collected from ULF.There was an increase in the transcript abundance of 20 miRNAs in the ULF EVs from the AI pregnant group,while 4 miRNAs had a lower abundance relative to the group not containing a conceptus.Additionally,4 miRNAs were more abun-dant in ULF EVs in the AI pregnant group relative to IVP-ET group(bta-mir-17,bta-mir-7-3,MIR7-1,MIR18A).Specific miRNAs in the ULF EVs were co-expressed with messenger RNAs expressed in extra-embryonic tissues and endome-trium,including genes that are known to be their targets.Conclusions The results provide biological insights into the participation of miRNAs in the regulation of trophoblast proliferation and differentiation,as well as in endometrium receptivity.The knowledge that in vitro cultured embryos can contribute to the altered abundance of specific miRNAs in the uterine lumen can lead to the development of cor-rective approaches to reduce conceptus losses during the first month of pregnancy in cattle.展开更多
Domoic acid(DA),a biotoxin,is produced by several species of marine dinoflagellate and diatom during harmful algal bloom events.DA is a neurotoxin,in humans and non-human primates,oral exposure to DA results in gastro...Domoic acid(DA),a biotoxin,is produced by several species of marine dinoflagellate and diatom during harmful algal bloom events.DA is a neurotoxin,in humans and non-human primates,oral exposure to DA results in gastrointestinal effects,while DA at higher doses leads to neurological symptoms,seizures and memory deficiency.Exposure of humans to DA occurs mainly through consumption of contaminated seafoods containing an accumulation of the toxin.Previously,it was unclear if DA can have toxic effects on the retina.We assessed the toxicity of DA in human retinal cells(ARPE-19)and in zebrafish embryos.DA significantly lowered ARPE-19 cell viability dose-dependently,and decreased anti-oxidative capacity,increased inflammation,and promoted cell death,possibly through modulating the NRF2 and NF-κB signalling pathways.Zebrafish embryos exposed to DA for 4 days from 1 day post fertilization(dpf)had an increase in mortality and a decrease in both hatching and heartbeat rate and exhibited morphological abnormalities.DA treatment also significantly downregulated expression of antioxidant genes and upregulated expression of inflammation mediators,as well as causing photoreceptor death in zebrafish embryos.The results demonstrate that consuming seafood containing DA will have potential toxic effects in human retinas.展开更多
Objective:To evaluate the effect of glutathione(GSH)supplementation in maturation and adaptation media on oocyte development,embryo quality,and oocyte viability after vitrification.Methods:The GSH concentrations were ...Objective:To evaluate the effect of glutathione(GSH)supplementation in maturation and adaptation media on oocyte development,embryo quality,and oocyte viability after vitrification.Methods:The GSH concentrations were classified into four groups(0,0.5,1.0,and 1.5 mM)which were added to the maturation medium.The maturation process was conducted in vitro for 24 h.Following maturation,oocytes were fertilized with Bali bull semen for 5-6 h and then cultured for 48 h.The morphological quality of ocytes matured with GSH addition and the vitrification method used was evaluated.Parameters assessed included maturation rate,fertilization rate,embryo development,post-vitrification oocyte morphology,and quality of post-vitrification oocytes with GSH added to the adaptation medium.Results:The addition of GSH to the maturation medium significantly improved oocyte quality and embryo development(P<0.05).Specifically,adding 1.5 mM GSH increased the percentage of oocytes reaching metaphase栻from 57.6%without GSH oocytes to 79.0%with 1.5 mM GSH,two-pronuclei fertilization from 47.0%to 72.7%,embryo development from 37.1%to 57.2%,morula formation from 14.6%to 33.7%,and blastocyst formation from 8.1%to 23.8%.Additionally,the survival rate of oocytes post-vitrification increased to 75%with GSH supplementation.Conclusions:The addition of 0.5-1.5 mM of GSH to the maturation and adaptation media significantly enhanced the metaphase栻stage,fertilization rates,cleavage rates,and the survival of oocytes after vitrification.Among the concentrations of 1.5 mM was the most effective in increasing oocyte development and maintaining oocytes viability post-vitrification.展开更多
Background:Polygonum multiflorum-induced liver injury(PM-DILI)has significantly hindered its clinical application and development.Methods:This study investigates the variation in content and toxicity of dian-thrones,t...Background:Polygonum multiflorum-induced liver injury(PM-DILI)has significantly hindered its clinical application and development.Methods:This study investigates the variation in content and toxicity of dian-thrones,the toxic components of P.multiflorum,during different processing cycles.We employed the ultra-high-performance liquid chromatography triple quadrupole mass spectrometry method to quantify six dianthrones in raw P.multiflorum and formulations processed with a method called nine cycles of steaming and sunning.Additionally,toxicity assessments were conducted using human normal liver cell line L02 and zebrafish embryos.Results:Results indicate a gradual reduction in dianthrones content with increasing processing cycles.Processed formulations exhibited significantly reduced cytotoxic-ity in L02 cells and hepatotoxicity in zebrafish embryos.Conclusions:Our findings elucidate the relationship between processing cycles and P.multiflorum toxicity,providing theoretical support for its safe use.展开更多
基金supported by Research on Breeding and Healthy Breeding Technology of Xueyu White Chicken(mating line)in Tibet Science and Technology Program(XZ202101ZY0002N)the National Key R&D Program Project(2022YFD1600902-4)Sichuan Province Regional Innovation Cooperation Project(2023YFQ0050)。
文摘Bacterial infections of avian embryos can lead to an increase in embryo mortality,and the proliferation of antimicrobial-resistant bacteria aggravates the situation.A low hatching rate also poses a challenge to the population of artificially bred Crested Ibises(Nipponia nippon).This study aims to determine the potential association between bacterial infection and the death of Crested Ibis embryos,and whether there is convergence between antimicrobial resistance and virulence in strain.In this study,13 Escherichia coli and 12 Proteus mirabilis isolates were recovered from dead Crested Ibis embryos.The pathogenicity examination confirmed the pathogenicity of all isolates,and multiple virulence genes detected by PCR-sequencing demonstrated the presence of irp2 and iuc D(100%),fim C and iss(92.31%)in E.coli,and uca A(58.33%)in P.mirabilis.Antimicrobial susceptibility test demonstrated that isolates were mainly resistant to amoxicillin(E.coli:76.92%,P.mirabilis:91.67%),cefazolin(E.coli:76.92%,P.mirabilis:91.67%),oxytetracycline(E.coli:92.31%,P.mirabilis:75.00%)and sulfamethoxazole-trimethoprim(E.coli:53.85%,P.mirabilis:33.33%),and more than 30%of isolates showed multidrug-resistance(MDR).Further analyses detected extended-spectrumβ-lactamase(ESBL)genes,of which blaTEM-1(E.coli:100%,P.mirabilis:100%)had the highest frequency,followed by the blaCTX-M-55(E.coli:92.31%,P.mirabilis:50%),blaCTX-M-14(E.coli:76.92%,P.mirabilis:33.33%),blaCTX-M-65(E.coli:15.38%,P.mirabilis:16.67%),and all isolates were negative for blaSHV and blaOXA.Pearson's correlation analysis showed a positive correlation between the presence ofβ-lactam resistance and ESBL genes,while mainly negative correlations were observed between the presence of ESBL genes and virulence genes.Furthermore,the conjugation experiment and PFGE revealed that the isolates were primarily polyclonal,and there was horizontal transfer of resistance or virulence genes by plasmids.Based on the results,E.coli and P.mirabilis were responsible for embryonic mortality of the ibises in this study.The co-presence and co-transfer of ESBL genes and virulence genes can pose a potential threat to the health of the Crested Ibis,and measures such as prudent use of antimicrobials,and constant surveillance of resistance and pathogenicity,must be implemented at the Crested Ibis breeding base.
基金supported by the National Natural Science Foundation of China(31872066 and 32272663)the Science and Technology Planning Project of Guangzhou,China(2023B01J2002)+2 种基金the Key Research and Development Program of Hainan,China(ZDYF2023XDNY052)the Seed Industry Engineering Project of Department of Agriculture and Rural Affairs of Guangdong,China(2022-NPY-00-004 and 2022-NBH00-001)the Litchi Industry Science and Technology Special Mission of Yunnan,China(202204BI090021)。
文摘Litchi has great economic significance as a global fruit crop.However,the advancement of litchi functional genomics has encountered substantial obstacles due to its recalcitrance to stable transformation.Here,we present an efficacious Agrobacterium tumefaciens-mediated transformation system in somatic embryos of‘Heiye'litchi.This system was developed through the optimization of key variables encompassing explant selection,A.tumefaciens strain delineation,bacterium concentration,infection duration,and infection methodology.The subsequent validation of the transformation technique in litchi was realized through the ectopic expression of LcMYB1,resulting in the generation of transgenic calli.However,the differentiation of transgenic calli into somatic embryos encountered substantial challenges.To delineate the intricate molecular underpinnings of LcMYB1's inhibitory role in somatic embryo induction,a comprehensive transcriptome analysis was conducted that encompassed embryogenic calli(C),globular embryos(G),and transgenic calli(TC).A total of 1,166 common differentially expressed genes(DEGs)were identified between C-vs.-G and C-vs.-TC.Gene Ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis revealed that these common DEGs were mostly related to plant hormone signal transduction pathways.Furthermore,RT-qPCR corroborated the pronounced down-regulation of numerous genes that are associated with somatic embryo induction within the transgenic calli.The development of this transformation system provides valuable support for functional genomics research in litchi.
基金This work was supported by the Science and Technology Development Plan Project of Jilin Province,China(20200402115NC).
文摘Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos.
基金supported by the Ministry of Science and Technology of China(Nos.2021YFA1101300 and 2020YFA0112500)the National Key R&D Program of China(2018YFA0801000)the National Natural Science Foundation of China(No.32170853)。
文摘As an increasingly used alternative to perfluorooctanoic acid(PFOA),hexafluoropropylene oxide trimer acid(HFPO-TA)has been widely detected in global water environments.However,little is known regarding its toxic effects on cardiovascular development.Here,zebrafish embryos were treated with egg water containing 0,60,120,or 240 mg/L HFPO-TA.Results showed that HFPO-TA treatment led to a significant reduction in both larval survival percentage and heart rate.Furthermore,HFPO-TA exposure caused severe pericardial edema and elongation of the sinus venous to bulbus arteriosus distance(SV-BA)in Tg(myl7:GFP)transgenic larvae,disrupting the expression of genes involved in heart development and thus causing abnormal heart looping.Obvious sprouting angiogenesis was observed in the 120 and 240 mg/L exposed Tg(fli:GFP)transgenic larvae.HFPO-TA treatment also impacted the mRNA levels of genes involved in the vascular endothelial growth factor(VEGF)pathway and embryonic vascular development.HFPO-TA exposure significantly decreased erythrocyte number in Tg(gata1:DsRed)transgenic embryos and influenced gene expression associated with the heme metabolism pathway.HFPO-TA also induced oxidative stress and altered the transcriptional levels of genes related to cell cycle and apoptosis,inhibiting cell proliferation while promoting apoptosis.Therefore,HFPO-TA exposure may induce abnormal development of the cardiovascular and hematopoietic systems in zebrafish embryos,suggesting it may not be a suitable or safe alternative for PFOA.
基金Project supported by the National Natural Science Foundation of China (Grant No.11974184)。
文摘The thermal-elastic martensitic transformation from high-temperature Ni_(2)In-type hexagonal structure to low-temperature TiNiSi-type orthorhombic structure has been widely studied in MnMX(M=Ni or Co,and X=Ge or Si)alloys.However,the answer to how the orthorhombic martensite nucleates and grows within the hexagonal parent is still unclear.In this work,the hexagonal-orthorhombic martensitic transformation in a Co and Ge co-substituted MnNiSi is investigated.One can find some orthorhombic laths embedded in the hexagonal parent at a temperature above the martensitic transformation start temperature(M_(s)).With the the sample cooing to M_(s),the laths turn broader,indicating that the martensitic transformation starts from these pre-existing orthorhombic laths.Microstructure observation suggests that these pre-existing orthorhombic laths do not originate from the hexagonal-orthorhombic martensitic transformation because of the difference between atomic occupations of doping elements in the hexagonal parent and those in the preexisting orthorhombic laths.The phenomenological crystallographic theory and experimental investigations prove that the pre-existing orthorhombic lath and generated orthorhombic martensite have the same crystallography relationship to the hexagonal parent.Therefore,the orthorhombic martensite can take these pre-existing laths as embryos and grow up.This work implies that the martensitic transformation in MnNiSi_(1-x)(CoNiGe)_(x) alloy is initiated by orthorhombic embryos.
基金Supported by National Natural Science Foundation of China(31070224)National Natural Science Foundation of China(30970219)Key Project from Science and Technology Department in Jilin Province(20080203)~~
文摘[Objective]This study was to screen out suitable genotypes and basic medium for the culture of maize mature embryos.[Method]Using mature embryos of nine maize genotypes as explants,the effects of genotypes and basic medium on callus induction and subculture were investigated.[Result]The genotypes performed better in callus induction and subculture were found in turn 853-35,853-209,Dan 34 and 81162.MS medium is better than N6 medium in the callus induction from maize embryos,while N6 medium is more suitable for callus subculture.[Conclusion]Our study further improved the tissue culture system in maize with mature embryos as explants.
基金Supported by Heilongjiang August First Land Reclamation University(Establishment of fast and high-frequency regeneration system of maize)~~
文摘In this study we studied the factors influencing the callus induction from mature embryos of maize inbred lines Qi 319, Zhen 58, Chang 7 -2, Lx 9801 and 81162, such as genotype, combination of plant growth regulators, and low-temperature pretreatment. The results showed that the induction rate of Qi 319 was the highest among the four genotypes tested; combination of 4.0 mg/L 2,4-D ＋ 0.5 mg/L 6-BA was suitable for inducing callus from mature embryos; three days of 4℃ pretreatment can promote the callus induction significantly. The indices optimized in the present study are helpful for establishing genetic transformation system in maize without considering seasonal variation.
文摘Fluctuations of levels of several endogenous plant hormones in isolated rice ( Oryza sativa ssp. japonica) embryos during early and mid-embryogenesis and early stages of germination were studied by enzyme-linked immunosorbent assay (ELISA). Embryos were collected at different days after pollination (DAP) and different days after imbibition (DAI) of mature seeds. The contents of gibberellin(1) (GA(1)), abscisic acid (ABA), isopentenyladenine and isopentenyladenosine ( iPAs), zeatin and zeatin riboside ( ZRs) were immunochemically assayed. The GA(1) level was the highest among all hormones tested. The variations of GA(1) levels were opposite with the ABA levels, with some exceptions. During early and mid-embryogenesis, the levels of GA(1) and ABA were the highest at 4 DAP. From 8 to 18 DAP, GA(1) level declined, whereas the ABA level increased. During germination, GA(1) level increased at 2 DAI whereas simultaneously the ABA content decreased. The highest ratio of GA(1)/ABA was observed at 2 DAI The levels of iPAs and ZRs were maxima in the embryos at 4 DAP, decreased to a very low level and maintained constant thereafter. Our results provide further evidence that GA(1) plays an important role in the early stages of embryo development and germination. The balance between GA(1) and ABA, rather than their absolute contents, controls these processes throughout the development, whereas iPAs and ZRs may play important roles in early embryogenesis. The use of isolated embryos as starting material avoids the usual interferences with other tissues such as the endosperm. In addition, this is the first report dealing with the hormonal balance of early-embryos in rice.
文摘An in situ hybridization technique for localization of calmodulin(CaM) mRNA in isolated entire embryo sacs and proembryos in Nicotiana tabacum L.cv.W38 has been developed. This technique can be applied to small amounts of materials in which a whole view of CaM mRNA distribution can be obtained. The authors revealed that CaM mRNA expression changes dramatically before and after fertilization. Especially interesting is that a prominent CaM mRNA band appears between the egg apparatus and polar nuclei temporarily during the period of pollination and fertilization. The band disappears just prior to fertilization and expands to a fan_shaped region that occupies the micropylar portion of the embryo sac. After fertilization, CaM mRNA accumulates in the elongated zygotes with higher concentration in their chalazal portion than in the micropylar portion. Such an asymmetrical pattern continues to manifest in the early proembryos. It is supposed that CaM mRNA may be involved in the early events and signaling steps associated with double fertilization and zygote polarization in higher plants.
文摘In ecological zone of Chengdu, Sichuan, microspore culture was carried out in Brassica napus L. to study the influencing factors on microspore culture. The results showed that the temperature on microspore formation stage, day and night temperature, disinfection solution of buds, cultivation concentration on microspore and strain-age were both important influencing factors on microspore culture. At a temperature below 5 ℃ or above 20 ℃, the material had a much lower embryo producing rate or even could not produce any embryo, but at the optimum temperature of 10 -15 ℃ the embryo yield was up to 300 pieces per bud; the best cultivation effect appeared when 0. 1% HgCl2 was used for disin- fection; the best density of microspore was 3 -4 buds per dish; In 2009, while strain-age was from 125 d to 150 d, the microspore embryo yield increased as strain-age increased. When stain-age was 150 days, the microspore embryo yield was up to the highest, but the yield declined after 150 days.
基金supported by the Institute for Basic Science (IBS-R022-D1)Global Learning&Academic Research Institution for Master’s/Ph D students and Post-Doc Program of the National Research Foundation of Korea Grant funded by the Ministry of Education (RS-2023-00301938)+1 种基金National Research Foundation of Korea Grant funded by the Korean government (RS-2024-00406152,MSIT)Additional financial support was provided by the 2024 Post-Doc Development Program of Pusan National University,Korea Medical Institute,and KREONET。
文摘Polystyrene nanoparticles pose significant toxicological risks to aquatic ecosystems,yet their impact on zebrafish(Danio rerio)embryonic development,particularly erythropoiesis,remains underexplored.This study used single-cell RNA sequencing to comprehensively evaluate the effects of polystyrene nanoparticle exposure on erythropoiesis in zebrafish embryos.In vivo validation experiments corroborated the transcriptomic findings,revealing that polystyrene nanoparticle exposure disrupted erythrocyte differentiation,as evidenced by the decrease in mature erythrocytes and concomitant increase in immature erythrocytes.Additionally,impaired heme synthesis further contributed to the diminished erythrocyte population.These findings underscore the toxic effects of polystyrene nanoparticles on hematopoietic processes,highlighting their potential to compromise organismal health in aquatic environments.
基金supported by the China Agriculture Research System(CARS-35-PIG)the National Natural Science Foundation of China(32272847,U22A20516)+1 种基金the National Key Research and Development Program of China(No.2021YFF1000602)the Key Research and Development Program of Shaanxi Province(No.2022ZDLNY01–04)。
文摘Background Increased backfat thickness of sows in early gestation is negative to reproductive performance.Endometrial receptivity is an important determinant of reproductive success,but it is unclear whether the effect of sow backfat thickness on litter size is associated with endometrial receptivity and whether melatonin treatment may have benefits.The present study seeks to answer these questions through in vitro and in vivo investigations.Results Excessive lipid deposition and lower melatonin levels in the uterus are detrimental to endometrial receptivity and embryo implantation in high backfat thickness sows.In cells treated with melatonin,the MT2/PI3K/LIF axis played a role in reducing lipid accumulation in porcine endometrial epithelium cells and improved endometrial receptivity.Furthermore,we found a reduction of lipids in the uterus after eight weeks of intraperitoneal administration of melatonin to HFD mice.Notably,melatonin treatment caused a significant reduction in the deposition of endometrial collagen,an increase in the number of glands,and repair of the pinopode structure,ultimately improving endometrial receptivity,promoting embryo implantation,and increasing the number of litter size of mice.Conclusions Collectively,the finding reveals the harmful effects of high backfat thickness sows on embryo implantation and highlight the role of melatonin and the MT2/PI3K/LIF axis in improving endometrial receptivity by enhancing metabolism and reducing the levels of uterine lipids in obese animals.
基金Support by Major Science and Technology Projects of Guangdong Province (2004A20102002)~~
文摘[Objective]The aim was to study the effective factors on culturing Prunus salicina cv.Zaoshi embryos in vitro.[Method]Different age,culture medium and GA3 which affected culturing of prumssalicina embryos were discussed.[Result]The result showed that on the part of 26-41 d,after the blooming period,the embryos remained tiny and retained endosperms and showed no signs of change after having cultured for three generations.On the part of 48 d,after the blooming period,the endosperms had disappeared,the embryos kept growing until they filled the seed cavity;when they were planted on the MS culture medium,their survival rate reached 77%,in its first generation,the response of embryos was discernible.On the part of 65 d,after the blooming period,4.5% of their embryos grew into shoots on the MS culture medium;with the age of embryos growing,the survival rate of shoots increased until it reached 26% when the fruits went into ripeness;the embryos produced calli in their first generation of culturing.On the part of 65-83 d,after the blooming period,the embryos produced calli through more than 2-3 generations.On the part of 88 d,after the blooming period,the survival rate of shoots on the WPM culturing base doubled compared with that on the MS culturing base;on the same culture medium,the embryos were inhibited from growing into shoots when BA,KT or 2,4-D was added on to the culture medium.The survival rate of shoots was increased remarkably when the seeds were treated in 1 000 mg/L GA3.[Conclusion] This study provided experimental basis for the establishment of Prunus salicina cv.Zaoshi,embryos rescue techniques and cross breading.
基金Supported by Natural Science Foundation (No. 30770124)SRTP Fund from Henan University of Science and Technology~~
文摘[Objective] The aim of this study was to manually isolate the microstructures such as ovule and embryo of Capsella bursa-pastoris (L.) Medic. by the non-enzymatic method. [Method] With the gum block and dissecting microscope, its floral characteristics, ovule and embryo were manually isolated and observed by the non-enzymatic method. [Result] C. bursa-pastoris had four calyces, four petals, tetradynamous stamen and silicle, while its embryogenesis contained globular embryo, heart-shaped embryo and torpedo-shaped embryo. [Conclusion] The microstructures such as ovule and embryo of plants become easier to be isolated manually with the viscosity of gum block, which has small damage on embryo or disadvantages of keeping original shapes, and also can used in multi fields studies. This test can also be accomplished in labs with poor experiment facilities.
基金Supported by Natural Science Foundation of Guangxi Province(Guikezi 0991096)~~
文摘[Objective] The aim was to explore the conditions of high frequency induction of embryonic callus and plant regeneration of maize. [Method] Immature embryos of maize inbred lines were used as explants to study the effect of different 2,4-D concentrations on the induction of callus,the effect of different 6-BA concentrations on the differentiation of test-tube plantlet,as well as the effect of different IBA concentrations on the rooting of test-tube plantlet. [Result] 2,4-D showed obvious effect on the induction of inducement rate of maize,and the optimum induction medium was：N6 ＋ 2 mg/L of 2,4-D ＋ 500 mg/L of CH ＋ 500 mg/L of Pro ＋10 mg/L of AgNO3; the optimum differentiation medium was：N6 ＋ 0.5 mg/L of BA ＋ 500 mg/L of Pro; the optimum for the rooting of test-tube plantlet was 1/2 MS ＋ 0.5 mg/L of IBA. [Conclusion] The study had provided basis for the genetic transformation of maize.
文摘The rapidly evolving environment of assisted reproductive technology(ART)requires consideration of how new innovations are reshaping clinical practice as much as research.In particular,there are three key areas that,while full of promise,also present significant challenges:the use of artificial intelligence(AI)in embryo selection,the impact of personalized medicine on ART success rates,and the ethical considerations of genetic screening of embryos[1].This letter is meant to provoke further discussion and highlight the need for harmonized global guidelines as these advances continue to reshape the reproductive medicine environment.
基金Agriculture and Food Research Initiative Competitive Grant no.2018-67015-31936 from the USDA National Institute of Food and Agriculture.
文摘Background MicroRNAs(miRNAs)are presented in the uterine lumen of many mammals,and in vitro experiments have determined that several miRNAs are important for the regulation of endometrial and trophoblast functions.Our aim was to identify and contrast the miRNAs present in extracellular vesicles(EVs)in the uterine lumen fluid(ULF)at the onset of attachment in cattle pregnancies(gestation d 18)initiated by artificial insemination(AI)or by the trans-fer of an in vitro-produced blastocyst(IVP-ET).A third group had no conceptus after the transfer of an IVP embryo.Results The abundance of 263 annotated miRNAs was quantified in the EVs collected from ULF.There was an increase in the transcript abundance of 20 miRNAs in the ULF EVs from the AI pregnant group,while 4 miRNAs had a lower abundance relative to the group not containing a conceptus.Additionally,4 miRNAs were more abun-dant in ULF EVs in the AI pregnant group relative to IVP-ET group(bta-mir-17,bta-mir-7-3,MIR7-1,MIR18A).Specific miRNAs in the ULF EVs were co-expressed with messenger RNAs expressed in extra-embryonic tissues and endome-trium,including genes that are known to be their targets.Conclusions The results provide biological insights into the participation of miRNAs in the regulation of trophoblast proliferation and differentiation,as well as in endometrium receptivity.The knowledge that in vitro cultured embryos can contribute to the altered abundance of specific miRNAs in the uterine lumen can lead to the development of cor-rective approaches to reduce conceptus losses during the first month of pregnancy in cattle.
基金funded by Namibe UniversityInstituto Nacional de Gestao de Bolsas de Estudo/Angola government through a PhD studentship program+1 种基金the TENOVUS Scotland(S20-02 to Xinhua Shu,the Chief Scientist Office/the RS Macdonald Charitable Trust(SNRF2021 to Xinhua Shu)the Lotus Scholarship Program of Hunan Province,China(2019-23 to Xinhua Shu)。
文摘Domoic acid(DA),a biotoxin,is produced by several species of marine dinoflagellate and diatom during harmful algal bloom events.DA is a neurotoxin,in humans and non-human primates,oral exposure to DA results in gastrointestinal effects,while DA at higher doses leads to neurological symptoms,seizures and memory deficiency.Exposure of humans to DA occurs mainly through consumption of contaminated seafoods containing an accumulation of the toxin.Previously,it was unclear if DA can have toxic effects on the retina.We assessed the toxicity of DA in human retinal cells(ARPE-19)and in zebrafish embryos.DA significantly lowered ARPE-19 cell viability dose-dependently,and decreased anti-oxidative capacity,increased inflammation,and promoted cell death,possibly through modulating the NRF2 and NF-κB signalling pathways.Zebrafish embryos exposed to DA for 4 days from 1 day post fertilization(dpf)had an increase in mortality and a decrease in both hatching and heartbeat rate and exhibited morphological abnormalities.DA treatment also significantly downregulated expression of antioxidant genes and upregulated expression of inflammation mediators,as well as causing photoreceptor death in zebrafish embryos.The results demonstrate that consuming seafood containing DA will have potential toxic effects in human retinas.
基金This study was financially supported by Universitas Hasanuddin through Penelitian Fundamental Kolaboratif(PFK)(letter of appointment number:00323/UN4.22/PT.0103/2023).
文摘Objective:To evaluate the effect of glutathione(GSH)supplementation in maturation and adaptation media on oocyte development,embryo quality,and oocyte viability after vitrification.Methods:The GSH concentrations were classified into four groups(0,0.5,1.0,and 1.5 mM)which were added to the maturation medium.The maturation process was conducted in vitro for 24 h.Following maturation,oocytes were fertilized with Bali bull semen for 5-6 h and then cultured for 48 h.The morphological quality of ocytes matured with GSH addition and the vitrification method used was evaluated.Parameters assessed included maturation rate,fertilization rate,embryo development,post-vitrification oocyte morphology,and quality of post-vitrification oocytes with GSH added to the adaptation medium.Results:The addition of GSH to the maturation medium significantly improved oocyte quality and embryo development(P<0.05).Specifically,adding 1.5 mM GSH increased the percentage of oocytes reaching metaphase栻from 57.6%without GSH oocytes to 79.0%with 1.5 mM GSH,two-pronuclei fertilization from 47.0%to 72.7%,embryo development from 37.1%to 57.2%,morula formation from 14.6%to 33.7%,and blastocyst formation from 8.1%to 23.8%.Additionally,the survival rate of oocytes post-vitrification increased to 75%with GSH supplementation.Conclusions:The addition of 0.5-1.5 mM of GSH to the maturation and adaptation media significantly enhanced the metaphase栻stage,fertilization rates,cleavage rates,and the survival of oocytes after vitrification.Among the concentrations of 1.5 mM was the most effective in increasing oocyte development and maintaining oocytes viability post-vitrification.
基金National Natural Science Foundation of China,Grant/Award Number:81973476Chinese Society of Toxicology,Grant/Award Number:CST2021CT101。
文摘Background:Polygonum multiflorum-induced liver injury(PM-DILI)has significantly hindered its clinical application and development.Methods:This study investigates the variation in content and toxicity of dian-thrones,the toxic components of P.multiflorum,during different processing cycles.We employed the ultra-high-performance liquid chromatography triple quadrupole mass spectrometry method to quantify six dianthrones in raw P.multiflorum and formulations processed with a method called nine cycles of steaming and sunning.Additionally,toxicity assessments were conducted using human normal liver cell line L02 and zebrafish embryos.Results:Results indicate a gradual reduction in dianthrones content with increasing processing cycles.Processed formulations exhibited significantly reduced cytotoxic-ity in L02 cells and hepatotoxicity in zebrafish embryos.Conclusions:Our findings elucidate the relationship between processing cycles and P.multiflorum toxicity,providing theoretical support for its safe use.
