The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose⁃catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monit...The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose⁃catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16℃in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni⁃NTA affinity column.Size⁃exclusion chromatography and SDS⁃PAGE analysis demonstrated that the puri⁃fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD^(+)as a coenzyme to NADP^(+).The optimal temperature and pH of the TeXDH were 40℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26%of the initial enzyme activity,while the divalent metal ions Mg^(2+)or Ca^(2+)could recover the enzyme activity of TeXDH,reaching 103.32%and 110.69%of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn^(2+),Ni^(2+),Cu^(2+),Zn^(2+),Co^(2+),and Cd^(2+)significantly inhibited the activity of TeXDH.ICP⁃MS and molecular doc⁃king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn^(2+)ions and 1 mol/L Mg^(2+)ion.Further⁃more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.展开更多
4-Nonylphenol(NP)is a kind of estrogen belonging to the endocrine disrupter,widely used in various agricultural and industrial goods.However,extensive use of NP with direct release to environment poses high risks to b...4-Nonylphenol(NP)is a kind of estrogen belonging to the endocrine disrupter,widely used in various agricultural and industrial goods.However,extensive use of NP with direct release to environment poses high risks to both human health and ecosystems.Herein,for the first time,we developed near-infrared(NIR)responsive upconversion luminescence nanosensor for NP detection.The Förster resonance energy transfer based upconversion nanoparticles(UCNPs)-graphene oxide sensor offers highly selective and sensitive detection of NP in linear ranges of 5−200 ng/mL and 200−1000 ng/mL under 980 nm and 808 nm excitation,respectively,with LOD at 4.2 ng/mL.The sensors were successfully tested for NP detection in real liquid milk samples with excellent recovery results.The rare-earth fluoride based upconversion luminescence nanosensor with NIR excitation wavelength,holds promise for sensing food,environmental,and biological samples due to their high sensitivity,specific recognition,low LOD,negligible autofluorescence,along with the deep penetration of NIR excitation sources.展开更多
Nicotine,also known as nicotinic norephedrine,is one of the main alkaloids present in tobacco plants.In recent years,due to the increase in tobacco production and smoking population,the environmental and health issues...Nicotine,also known as nicotinic norephedrine,is one of the main alkaloids present in tobacco plants.In recent years,due to the increase in tobacco production and smoking population,the environmental and health issues caused by nicotine have become increasingly severe.Traditional methods have proven ineffective in efficiently degrading residual nicotine.To address this issue,scientists both domestically and internationally have turned to biodegradation methods to tackle the environmental and health problems caused by residual nicotine.In this study,an enrichment method was used to screen bacteria with nicotine-degrading capabilities from the soil of tobacco planting sites at the Tobacco Research Institute of Heilongjiang in Bin County,Harbin City.Through phenotypic observations and 16S rDNA identification,a bacterial strain identified as Pseudomonas hunanensis MGJ-2 was isolated,capable of utilizing nicotine as a carbon and nitrogen source for growth.High-performance liquid chromatography(HPLC)-1 analysis revealed that within 25 h,strain MGJ-2 could degrade nicotine 500 mg·L^(-1) with an efficiency exceeding 99.9%.Strain MGJ-2 was applied to tobacco,and after 15 days of incubation and fermentation,it degraded 10.57%of nicotine in tobacco.Overall,the discovery of strain MGJ-2 enriched the resources of nicotine-degrading strains.Its remarkable biodegradation performance held immense potential for future biodegradation of nicotine in tobacco.展开更多
In order to elucidate the relationship between the pathogen carriage rate in seeds of muskmelon(Cucumis melo L.)and the incidence rate of Fusarium wilt of muskmelon(FWM),as well as to identify potential biological con...In order to elucidate the relationship between the pathogen carriage rate in seeds of muskmelon(Cucumis melo L.)and the incidence rate of Fusarium wilt of muskmelon(FWM),as well as to identify potential biological control agents against FWM,this study conducted both pot and field experiments to evaluate the efficacy of Gongzhulingmycin on FWM and its impact on muskmelon yield.The results indicated that the pathogen carriage rates of different species in muskmelon seeds varied significantly,showing a positive correlation with disease incidence during the seedling stage.The results from pot tests indicated that in comparison to the control,disease indices were significantly reduced following treatment with prochloraz and 100 times Gongzhulingmycin at both 7 days and 14 days post-emergence of FWM symptoms.Concurrently,root growth was enhanced.Field experiment outcomes demonstrated that relative to the control,there was a decrease in FWM incidence during the fruit-setting stage,along with an increase in theoretical output per square kilometer for muskmelon.Although the efficacy of Gongzhulingmycin against FWM was lower than that observed for prochloraz,it exhibited significant effects on biomass enhancement and disease resistance.Therefore,it showed promise as a potential biological control agent for managing FWM.展开更多
Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expres...Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro- MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly （P〈0.05）. After knocked down MSTN gene, P21 expression decreased （P〈0.05）, but CDK2 gene expression increased （P〈0.05）. These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.展开更多
For solving the dilemma of the short exothermic life-span of WO_(3)based metastable interstitial composites(MICs)with extensive application prospect,this paper has firstly designed the promising antiwetting Al/WO_(3)M...For solving the dilemma of the short exothermic life-span of WO_(3)based metastable interstitial composites(MICs)with extensive application prospect,this paper has firstly designed the promising antiwetting Al/WO_(3)MICs via electrophoresis assembly of nano-Al and WO_(3)particles fabricated by hydrothermal synthesis method,followed by the subsequent fluorination treatment.A combination of X ray diffraction(XRD),field emission scanning electron microscope(FESEM),energy dispersive X-ray spectroscopy(EDX),and Fourier transform infrared spectroscopy(FT-IR)techniques were utilized in order to characterize the crystal structure,microstructure,and elemental composition distribution of target films after different natural exposure tests.The product with uniform distribution and high purity possesses a high contact angle of~170°and a minute sliding angle of~1°,and displays the outstanding anti-wetting property using droplets with different surface tensions.It also shows great moisture stability in high relative-humidity circumstances after one year of the natural exposure experiment.Notably,the heat output of a fresh sample can reach up to 2.3 kJ/g and retain 96%after the whole exposure test,showing outstanding thermo-stability for at least one year.This work further proposed the mechanism of antiwetting Al/WO_(3)MICs considering the variation tendency of their DSC curve,providing a valuable theoretical reference for designing other self-protected MICs with a long exothermic life-span applied in wide fields of national defense,military industry,etc.展开更多
The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this ...The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this research field obtained over the past two years.This report summarizes the major progress of the basic researches about the origin of life in China during 2018–2020.展开更多
12%difenoconazole+fluxapyroxad SC(commercial name:Jiangong)was first released by BASF in China in 2016.It has been registered to control many diseases,including pear scab,apple Alternaria leaf spot,tomato early blight...12%difenoconazole+fluxapyroxad SC(commercial name:Jiangong)was first released by BASF in China in 2016.It has been registered to control many diseases,including pear scab,apple Alternaria leaf spot,tomato early blight,cucumber powdery mildew,etc.This study evaluated the bioactivity of Jiangong against Alternaria alternata and explored variations of phyllosphere microorganisms in both asymptomatic and tobacco brown spot leaves at different persistence periods(0,5,10,and 15 days post-fungicide application)using high-throughput sequencing technology.The results indicated that Jiangong effectively inhibited mycelial growth(average EC_(50) value of 0.51μg/mL),conidia germination(average EC_(50) value of 3.47μg/mL),and the carbon metabolism of A.alternata.Both asymptomatic and symptomatic leaves presented complex microbial communities.Higher fungal diversity was noted in asymptomatic leaves,while higher bacterial diversity was found in symptomatic leaves.After application,the diversity and abundance of microbial community structures in both types of leaves changed over time.Fungal microbiome communities showed greater sensitivity than bacterial groups,with the microbiome communities of asymptomatic leaves being more affected than those of symptomatic leaves.Fungal community diversity decreased for both symptomatic and asymptomatic leaves after 5 days of application,while the diversity of fungal community in symptomatic leaves showed an upward trend after 10 days of application.Meanwhile,bacterial community diversity increased in both symptomatic and asymptomatic leaves after 5 days of application but then declined in asymptomatic leaves after 15 days.The abundance of the dominant function group of phyllosphere bacteria(metabolism,genetic information processing,environmental information processing)was not affected by the application of Jiangong.However,the abundance of the dominant function group of phyllosphere fungi(animal pathogen-endophyte-wood saprotroph,endophyte-plant pathogen,plant pathogen-undefined saprotroph)was significantly affected by the application of Jiangong,and high variation was found in symptomatic leaves than that of asymptomatic leaves.The application of Jiangong-induced alterations in the community structure of the tobacco phyllosphere microbiome provides a basis for future tobacco brown spot control strategies based on phyllospheric microecology.展开更多
Objective In kinesin-3,the neck coil correlates with the following segments to form an extended neck that contains a characteristic hinge diverse from a proline in KIF13B to a long flexible linker in KIF1A.The functio...Objective In kinesin-3,the neck coil correlates with the following segments to form an extended neck that contains a characteristic hinge diverse from a proline in KIF13B to a long flexible linker in KIF1A.The function of this neck hinge for controlling processive movement,however,remains unclear.Methods We made a series of modifications to the neck hinges of KIF13B and KIF1A and tested their movement using a single-molecule motility assay.Results In KIF13B,the insertion of flexible residues before or after the proline differentially impacts the processivity or velocity,while the removal of this proline increases the both.In KIF1A,the deletion of entire flexible neck hinge merely enhances the processivity.The engineering of these hinge-truncated necks of kinesin-3 into kinesin-1 similarly boosts the processive movement of kinesin-1.Conclusion The neck hinge in kinesin-3 controls its processive movement and proper modifications tune the motor motility,which provides a novel strategy to reshape the processive movement of kinesin motors.展开更多
Objective Detection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability,health,and metabolic rate.After exposure to environmental stimuli,both the internal reference g...Objective Detection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability,health,and metabolic rate.After exposure to environmental stimuli,both the internal reference gene and target gene would be degraded.As a result,it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation.This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment.Methods The new RNA was labeled with 5-ethyluridine(5-EU)instead of uracil,and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of“Click Chemistry”and magnetic bead screening.Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon(M.B.)and quantitative reverse transcription PCR(qRT-PCR).Results The bacterial nascent RNA captured by“Click Chemistry”screening can be used as a reverse transcription template to form cDNA.Combined with the fluorescent molecular beacon M.B.1,the synthesis rate of rRNA at 37℃is 1.2 times higher than that at 15℃.The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃and 16℃when analyzed with nascent RNA rather than total RNA,enabling accurate detection of RNA transcription rates.Conclusion Compared to other article reported experimental methods that utilize screening magnetic columns,the technical scheme employed in this study is more suitable for bacteria,and the operation steps are simple and easy to implement,making it an effective RNA capture method for researchers.展开更多
Avian influenza virus is one of the main pathogens causing avian influenza.In this experiment,the avian influenza hemagglutinin(HA)gene was transferred into tobacco by Agrobacterium tumefaciens-mediated method using k...Avian influenza virus is one of the main pathogens causing avian influenza.In this experiment,the avian influenza hemagglutinin(HA)gene was transferred into tobacco by Agrobacterium tumefaciens-mediated method using kanamycin as a resistance marker to produce HA type edible vaccine designed for avian influenza hemagglutinin protein.The fusion of cholera toxin B subgene(CTB)and avian influenza HA was initiated by CaMV35S promoter,and then transformed into Nicotiana benthamiana to induce callus and adventitious bud differentiation,bud elongation,and growth and root induction.The total genomic DNA of 28 transgenic tobacco plants was extracted,and the CTB sequence of the CTB-HA fusion gene was used as a template to design primers for PCR amplification.Eight of them were positive,and four of them were expressed at the RNA level after RNA extraction and RT-PCR amplification.In western blot detection of protein extraction,two strains were shown at the position of the target protein.The results provided material support for the preparation of a transgenic plant oral vaccine against HA infection,and provided a theoretical basis for accelerating the development of a transgenic plant vaccine.展开更多
In order to optimize the ultrasonic extraction technique for the total flavonoid of leaf yellows plus, the contents of 21 leaf yellows plus total flavonoid from four regions in Heilongjiang Province were comparatively...In order to optimize the ultrasonic extraction technique for the total flavonoid of leaf yellows plus, the contents of 21 leaf yellows plus total flavonoid from four regions in Heilongjiang Province were comparatively analyzed. The ultrasonic extraction technology was optimized by Box-Behnken response surface method, and the total flavonoid content of 21 kinds of Acanthopanax senticosus(Rupr. et Maxim.) Harms from different producing areas were analyzed by cluster analysis. The optimal process conditions were determined as ultrasonic time 30 min, solid-liquid ratio 1 : 12 and ultrasonic power 250 W, and the average yield of the total flavonoid was 1.453 mg·g^ (-1). By optimizing the ultrasonic-assisted extraction method, the total flavonoid content from different producing areas was compared in the experiment, which provided certain data support for the optimization of the extraction process in the future and laid a certain theoretical foundation for the quality analysis of Chinese medicinal materials.展开更多
Background Aphis gossypii(Hemiptera:Aphididae)is a worldwide polyphagous phloem-feeding agricultural pest,and it can produce offspring by sexual or asexual reproduction.Compared with dozens of generations by parthenog...Background Aphis gossypii(Hemiptera:Aphididae)is a worldwide polyphagous phloem-feeding agricultural pest,and it can produce offspring by sexual or asexual reproduction.Compared with dozens of generations by parthenogenesis,sexual reproduction is performed in only one generation within one year,and little is known about the sexual reproduction of A.gossypii.In this study,sexual females of A.gossypii were successfully obtained through a previously established induction platform,and the morphological characteristics,developmental dynamics,and temporal gene expression were examined.Subsequently,signaling pathways potentially involved in regulating the growth,development,and reproduction of sexual females were investigated.Results The morphological observation showed that from the 1st instar nymph to adult,sexual females exhibited a gradually deepened body color,an enlarged body size,longer antennae with a blackened end,and obviously protruding cauda(in adulthood).The anatomy found that the ovaries of sexual females developed rapidly from the 2^(nd)instar nymph,and the embedded oocytes matured in adulthood.In addition,time-course transcriptome analysis revealed that gene expression profiles across the development of sexual females fell into 9 clusters with distinct patterns,in which gene expression levels in clusters 1,5,and 8 peaked at the 2^(nd)instar nymphal stage with the largest number of up-regulated genes,suggesting that the 2^(nd)instar nymph was an important ovary development period.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis revealed that a large number of genes in the sexual female adult were enriched in the TGF-beta signaling pathway and Forkhead box O(FoxO)signaling pathway,highlighting their important role in sexual female adult development and reproduction.Conclusion The morphological changes of the sexual female at each developmental stage were revealed for the first time.In addition,time-course transcriptomic analyses suggest genes enriched in the TGF-beta signaling pathway and FoxO signaling pathway probably contribute to regulating the development and oocyte maturation of sexual females.Overall,these findings will facilitate the regulating mechanism research in the growth and development of sexual females by providing candidate genes.展开更多
A novel 3D metal-organic framework(MOF)[Pr_(2)(L)_(3)(H_(2)O)5·H_(2)O]n(Pr-1),(H_(2)L=4,4'-oxybis(benzoic acid))with a rare structure of broken layer net,was constructed under the condition of solvothermal sy...A novel 3D metal-organic framework(MOF)[Pr_(2)(L)_(3)(H_(2)O)5·H_(2)O]n(Pr-1),(H_(2)L=4,4'-oxybis(benzoic acid))with a rare structure of broken layer net,was constructed under the condition of solvothermal synthesis.The struc-ture and crystal net were analyzed and characterized.This rod net of Pr-1 is new to both RCSR and ToposPro data-bases,and is named as rn-12 as suggested.Due to the luminescent properties of H_(2)L and Pr(Ⅲ),the solid-state fluo-rescence property and sensing performance(solvents and metal ions)of Pr-1 were investigated.The sensing experi-ments indicated that Pr-1 could act as a fluorescence sensor to detect Cd^(2+)ions with good sensitivity.In addition,antibacterial activities show that Pr-1 exhibited stronger antibacterial activity against Escherichia coli(E.coli),Staphylococcus aureus(S.aureus),and Bacillus subtilis(B.subtilis)compared to synthetic materials.展开更多
Objective Hepatocyte nuclear factor 4-alpha(HNF4A)is a critical transcription factor in the liver and pancreas.Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1(MODY1).Notably,MODY1 patients with HN...Objective Hepatocyte nuclear factor 4-alpha(HNF4A)is a critical transcription factor in the liver and pancreas.Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1(MODY1).Notably,MODY1 patients with HNF4A pathogenic mutations exhibit decreased responses to arginine and reduced plasma triglyceride levels,but the mechanisms remain unclear.This study aims to investigate the potential target genes transcriptionally regulated by HNF4A and explore its role in these metabolic pathways.Methods A stable 293T cell line expressing the HNF1A reporter was overexpressed with HNF4A.RNA sequencing(RNA-seq)was performed to analyze transcriptional differences.Transcription factor binding site prediction was then conducted to identify HNF4A binding motifs in the promoter regions of relevant target genes.Results RNA-seq results revealed a significant upregulation of transmembrane 4 L six family member 5(TM4SF5)mRNA in HNF4A-overexpressing cells.Transcription factor binding predictions suggested the presence of five potential HNF4A binding motifs in the TM4SF5 promoter.Finally,we confirmed that the DR1 site in the-57 to-48 region of the TM4SF5 promoter is the key binding motif for HNF4A.Conclusion This study identified TM4SF5 as a target gene of HNF4A and determined the key binding motif involved in its regulation.Given the role of TM4SF5 as an arginine sensor in mTOR signaling activation and triglyceride secretion,which closely aligns with phenotypes observed in MODY1 patients,our findings provide novel insights into the possible mechanisms by which HNF4A regulates triglyceride secretion in the liver and arginine-stimulated insulin secretion in the pancreas.展开更多
Objective The nucleolar protein PES1(Pescadillo homolog 1)plays critical roles in ribosome biogenesis and cell cycle regulation,yet its involvement in cellular senescence remains poorly understood.This study aimed to ...Objective The nucleolar protein PES1(Pescadillo homolog 1)plays critical roles in ribosome biogenesis and cell cycle regulation,yet its involvement in cellular senescence remains poorly understood.This study aimed to comprehensively investigate the functional consequences of PES1 suppression in cellular senescence and elucidate the molecular mechanisms underlying its regulatory role.Methods Initially,we assessed PES1 expression patterns in two distinct senescence models:replicative senescent mouse embryonic fibroblasts(MEFs)and doxorubicin-induced senescent human hepatocellular carcinoma HepG2 cells.Subsequently,PES1 expression was specifically downregulated using siRNA-mediated knockdown in these cell lines as well as additional relevant cell types.Cellular proliferation and senescence were assessed by EdU incorporation and SA-β-gal staining assays,respectively.The expression of senescence-associated proteins(p53,p21,and Rb)and SASP factors(IL-6,IL-1β,and IL-8)were analyzed by Western blot or qPCR.Furthermore,Northern blot and immunofluorescence were employed to evaluate pre-rRNA processing and nucleolar morphology.Results PES1 expression was significantly downregulated in senescent MEFs and HepG2 cells.PES1 knockdown resulted in decreased EdU-positive cells and increased SA-β-gal-positive cells,indicating proliferation inhibition and senescence induction.Mechanistically,PES1 suppression activated the p53-p21 pathway without affecting Rb expression,while upregulating IL-6,IL-1β,and IL-8 production.Notably,PES1 depletion impaired pre-rRNA maturation and induced nucleolar stress,as evidenced by aberrant nucleolar morphology.Conclusion Our findings demonstrate that PES1 deficiency triggers nucleolar stress and promotes p53-dependent(but Rb-independent)cellular senescence,highlighting its crucial role in maintaining nucleolar homeostasis and regulating senescence-associated pathways.展开更多
Esophageal cancer(EC)is one of the most common malignancies in the world,and there is no specific treatment drug for esophageal cancer yet.Doramectin(DRM)is a broad-spectrum anti-parasitic drug,and it plays an importa...Esophageal cancer(EC)is one of the most common malignancies in the world,and there is no specific treatment drug for esophageal cancer yet.Doramectin(DRM)is a broad-spectrum anti-parasitic drug,and it plays an important role in the treatment of animal diseases,while DRM has not been reported for the treatment of esophageal squamous cell carcinoma(ESCC).The purpose of this study was to investigate the anticancer effects and potential molecular mechanisms of DRM in ESCC.In the present study,the impact of DRM on the viability of ESCC was examined by methylthiazolyldiphenyl-tetrazolium bromide(MTT).Autophagy was measured by transmission electron microscopy(TEM),Western blot and immunohistochemistry.The apoptosis rate was measured by Western blot,flow cytometry and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL).Meanwhile,autophagy inhibition was achieved by using chloroquine(CQ).After autophagy inhibition,cell proliferation and cloning ability were significantly inhibited,and the expression level of apoptotic protein was significantly changed compared with that of DRM alone.Additionally,Eca109-derived xenografts were established for testing the DRM-induced autophagy in vivo.It was found that DRM significantly inhibited the proliferation of Eca109 and EC9706 cells in vitro and in vivo in a dose-dependent manner by activating autophagy.DRM was able to significantly repress colony formation in Eca109 and EC9706 cells in vitro.At the same time,DRM could induce apoptosis of ESCC in vitro,it was also regulated through mitochondrial pathways.Meanwhile,DRM induced autophagy and inhibited the proliferation of ESCC,and exhibited little toxicity in organs in vivo.Moreover,DRM-induced autophagy could inhibit the apoptosis of EC in vitro and in vivo.Further experiment suggested that DRM might induce autophagy by the Akt/mTOR pathway.In conclusion,the present study was the first to clarify that DRM could inhibit Eca109 and EC9706 cells proliferation through activating autophagy by the Akt/mTOR pathway.DRM might be a potentially effective treatment for EC.展开更多
Platycodon grandiflorum A.DC.(PAl)C)root was taken as experiment material to extract polysaccharide.On the base of single factor tests(extraction time,extraction temperature,liquid-solid ratio,solvent pH value and NaC...Platycodon grandiflorum A.DC.(PAl)C)root was taken as experiment material to extract polysaccharide.On the base of single factor tests(extraction time,extraction temperature,liquid-solid ratio,solvent pH value and NaCl concentration),the study concluded the main factors affecting the extraction of PADC polysaccharide,which are liquid-solid ratio,extraction time and extraction temperature.Then through central-composite test design,the extraction conditions were concluded as liquid-solid ratio 34.43,extraction time 89.83 min and extraction temperature 52.47℃.By means of validation experiments,the adequacy of this model was confirmed.展开更多
Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescen...Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescence material Ru(bpy)32+·6H2O, and then the particles were characterized via transmission electron microscope. The fluorescence nanoparticles were conjugated with plasmid DNA to form complexes, and then treated with ultrasound and DNase I. pEGAD plasmid DNA-nanoparticle complexes were co-cultured with plant suspension cells of Dioscrea Zigiberensis G H Wright, and treated with ultrasound. The results show that the diameter of the fluorescence starch-nanoparticles is 50-100 nm. DNA-nanoparticle complexes can protect DNA from ultrasound damage as well as from DNase I cleavage. Mediated by ultrasound, pEGAD plasmid DNA-nanoparticle complexes can pierce into the cell wall, cell membrane and nucleus membrane of plant suspension cells. The green fluorescence protein(GFP) gene at a high frequency exceeds 5%. This nano-biomaterial can efficiently solve the problem that exterior genes cannot traverse the plant cell wall easily.展开更多
Methionine （Met） and lysine （Lys） have been reported as the first two limiting amino acids （AA） for maximum milk yield and milk protein production. Supplying these AA may improve microbial protein synthesis and ...Methionine （Met） and lysine （Lys） have been reported as the first two limiting amino acids （AA） for maximum milk yield and milk protein production. Supplying these AA may improve microbial protein synthesis and therefore improve milk production without adding excess N to the environment. This observation utilized fermented soybean meal （SBM）, cottonseed meal （CSM）, rapeseed meal （RSM） and corn by Bacillus subtilis 168 and Leuconostoc mesenteroides as core feedstuffs to produce special biological protein feed for dairy cow. The results showed that the milk production, milk protein percentage, milk fat percentage and milk DM percentage of test groups in trial period were significantly more than those of the control group （P〈0.01）, the results showed that adding fermenting protein feed in dairy cow diets could significantly improve milk yield, milk protein and milk fat content. The economic benefits of actual application were analyzed, the group of 0.5% was the best compared with the other groups.展开更多
基金湖南省教育厅基金优秀青年项目(No.22B0482)湖南科技大学博士启动基金(No.E51992 and E51993)资助。
文摘The xylitol dehydrogenase(XDH)is a crucial enzyme involved in the xylose utilization in pentose⁃catabolizing yeasts and fungi.In addition to producing xylulose,XDH can also be employed to develop a biosensor for monitoring xylitol concentration.In this study,the gene encoding the thermophilic fungus Talaromyces emersonii XDH(TeXDH)was heterologously expressed in Escherichia coli BL21(DE3)at 16℃in the soluble form.Recombinant TeXDH with high purity was purified by using a Ni⁃NTA affinity column.Size⁃exclusion chromatography and SDS⁃PAGE analysis demonstrated that the puri⁃fied recombinant TeXDH exists as a native trimer with a molecular mass of approximately 116 kD,and is composed of three identical subunits,each with a molecular weight of around 39 kD.The TeXDH strictly preferred NAD^(+)as a coenzyme to NADP^(+).The optimal temperature and pH of the TeXDH were 40℃and 10.0,respectively.After EDTA treatment,the enzyme activity of TeXDH decreased to 43.26%of the initial enzyme activity,while the divalent metal ions Mg^(2+)or Ca^(2+)could recover the enzyme activity of TeXDH,reaching 103.32%and 110.69%of the initial enzyme activity,respectively,making them the optimal divalent metal ion cofactors for TeXDH enzyme.However,the divalent metal ions of Mn^(2+),Ni^(2+),Cu^(2+),Zn^(2+),Co^(2+),and Cd^(2+)significantly inhibited the activity of TeXDH.ICP⁃MS and molecular doc⁃king studies revealed that 1 mol/L of TeXDH bound 2 mol/L Zn^(2+)ions and 1 mol/L Mg^(2+)ion.Further⁃more,TeXDH exhibited a high specificity for xylitol,laying the foundation for the development of future xylitol biosensors.
文摘4-Nonylphenol(NP)is a kind of estrogen belonging to the endocrine disrupter,widely used in various agricultural and industrial goods.However,extensive use of NP with direct release to environment poses high risks to both human health and ecosystems.Herein,for the first time,we developed near-infrared(NIR)responsive upconversion luminescence nanosensor for NP detection.The Förster resonance energy transfer based upconversion nanoparticles(UCNPs)-graphene oxide sensor offers highly selective and sensitive detection of NP in linear ranges of 5−200 ng/mL and 200−1000 ng/mL under 980 nm and 808 nm excitation,respectively,with LOD at 4.2 ng/mL.The sensors were successfully tested for NP detection in real liquid milk samples with excellent recovery results.The rare-earth fluoride based upconversion luminescence nanosensor with NIR excitation wavelength,holds promise for sensing food,environmental,and biological samples due to their high sensitivity,specific recognition,low LOD,negligible autofluorescence,along with the deep penetration of NIR excitation sources.
基金Supported by the Research on Biofermentation Technology of Domestic Cigar Tobacco(202115010534-JS-178)(2021)。
文摘Nicotine,also known as nicotinic norephedrine,is one of the main alkaloids present in tobacco plants.In recent years,due to the increase in tobacco production and smoking population,the environmental and health issues caused by nicotine have become increasingly severe.Traditional methods have proven ineffective in efficiently degrading residual nicotine.To address this issue,scientists both domestically and internationally have turned to biodegradation methods to tackle the environmental and health problems caused by residual nicotine.In this study,an enrichment method was used to screen bacteria with nicotine-degrading capabilities from the soil of tobacco planting sites at the Tobacco Research Institute of Heilongjiang in Bin County,Harbin City.Through phenotypic observations and 16S rDNA identification,a bacterial strain identified as Pseudomonas hunanensis MGJ-2 was isolated,capable of utilizing nicotine as a carbon and nitrogen source for growth.High-performance liquid chromatography(HPLC)-1 analysis revealed that within 25 h,strain MGJ-2 could degrade nicotine 500 mg·L^(-1) with an efficiency exceeding 99.9%.Strain MGJ-2 was applied to tobacco,and after 15 days of incubation and fermentation,it degraded 10.57%of nicotine in tobacco.Overall,the discovery of strain MGJ-2 enriched the resources of nicotine-degrading strains.Its remarkable biodegradation performance held immense potential for future biodegradation of nicotine in tobacco.
基金Supported by the Key Research and Development Project of the Department of Science and Technology of Jilin Province(20230203175SF)。
文摘In order to elucidate the relationship between the pathogen carriage rate in seeds of muskmelon(Cucumis melo L.)and the incidence rate of Fusarium wilt of muskmelon(FWM),as well as to identify potential biological control agents against FWM,this study conducted both pot and field experiments to evaluate the efficacy of Gongzhulingmycin on FWM and its impact on muskmelon yield.The results indicated that the pathogen carriage rates of different species in muskmelon seeds varied significantly,showing a positive correlation with disease incidence during the seedling stage.The results from pot tests indicated that in comparison to the control,disease indices were significantly reduced following treatment with prochloraz and 100 times Gongzhulingmycin at both 7 days and 14 days post-emergence of FWM symptoms.Concurrently,root growth was enhanced.Field experiment outcomes demonstrated that relative to the control,there was a decrease in FWM incidence during the fruit-setting stage,along with an increase in theoretical output per square kilometer for muskmelon.Although the efficacy of Gongzhulingmycin against FWM was lower than that observed for prochloraz,it exhibited significant effects on biomass enhancement and disease resistance.Therefore,it showed promise as a potential biological control agent for managing FWM.
基金Supported by the Major Special Projects of New Product Training of Transgenic Organisms(zx080072008-2008)
文摘Myostatin （MSTN） is a negative regulator of skeletal muscle growth, in order to study the effect of inhibition MSTN expression on the proliferation of bovine skeletal muscle satellite cells, we constructed co-expression vector pcDNA3.1-Pro- MSTNshRNA, transfected it into muscle satellite cells by Liposome 2000, and detected cell proliferation changes by CCK-8 method and flow cytometry after 48 h. The expressions of P21 and CDK2 were detected by Western blot and real-time PCR. The results showed that the cell vitality of experimental groups significantly increased than that of the negative control, and cells in S phase also increased significantly （P〈0.05）. After knocked down MSTN gene, P21 expression decreased （P〈0.05）, but CDK2 gene expression increased （P〈0.05）. These results indicated that MSTN gene expression was associated with P21 and CDK2, the proliferation of skeletal muscle satellite cells could be promoted while MSTN was inhibited, which provided a theoretical basis for the study on transgenic cattle.
基金funded by the financial support from National Natural Science Foundation of China(Grant No 21805014 and No82102635)Science and Technology Research Project of Chongqing Education Board(Grant No.KJQN201901428)。
文摘For solving the dilemma of the short exothermic life-span of WO_(3)based metastable interstitial composites(MICs)with extensive application prospect,this paper has firstly designed the promising antiwetting Al/WO_(3)MICs via electrophoresis assembly of nano-Al and WO_(3)particles fabricated by hydrothermal synthesis method,followed by the subsequent fluorination treatment.A combination of X ray diffraction(XRD),field emission scanning electron microscope(FESEM),energy dispersive X-ray spectroscopy(EDX),and Fourier transform infrared spectroscopy(FT-IR)techniques were utilized in order to characterize the crystal structure,microstructure,and elemental composition distribution of target films after different natural exposure tests.The product with uniform distribution and high purity possesses a high contact angle of~170°and a minute sliding angle of~1°,and displays the outstanding anti-wetting property using droplets with different surface tensions.It also shows great moisture stability in high relative-humidity circumstances after one year of the natural exposure experiment.Notably,the heat output of a fresh sample can reach up to 2.3 kJ/g and retain 96%after the whole exposure test,showing outstanding thermo-stability for at least one year.This work further proposed the mechanism of antiwetting Al/WO_(3)MICs considering the variation tendency of their DSC curve,providing a valuable theoretical reference for designing other self-protected MICs with a long exothermic life-span applied in wide fields of national defense,military industry,etc.
文摘The development of Chinese space science and technology plays a great role in promoting the researches in the field of the origin of life.With the multidisciplinary cooperation,there are fruitful achievements in this research field obtained over the past two years.This report summarizes the major progress of the basic researches about the origin of life in China during 2018–2020.
基金Supported by China National Tobacco Corporation[No.110202101048(LS-08)]Hundred’Level Innovative Talent Foundation of Guizhou Province(No.GCC[2022]028-1,GCC[2023]108)+2 种基金Guizhou Science Technology Foundation(No.ZK[2021]Key036)the National Natural Science Foundation of China(No.32160522)Guizhou Province Applied Technology Research and Development Funding Post-subsidy Project and Guizhou Tobacco Company(No.2020XM03,2020XM22,2024XM06).
文摘12%difenoconazole+fluxapyroxad SC(commercial name:Jiangong)was first released by BASF in China in 2016.It has been registered to control many diseases,including pear scab,apple Alternaria leaf spot,tomato early blight,cucumber powdery mildew,etc.This study evaluated the bioactivity of Jiangong against Alternaria alternata and explored variations of phyllosphere microorganisms in both asymptomatic and tobacco brown spot leaves at different persistence periods(0,5,10,and 15 days post-fungicide application)using high-throughput sequencing technology.The results indicated that Jiangong effectively inhibited mycelial growth(average EC_(50) value of 0.51μg/mL),conidia germination(average EC_(50) value of 3.47μg/mL),and the carbon metabolism of A.alternata.Both asymptomatic and symptomatic leaves presented complex microbial communities.Higher fungal diversity was noted in asymptomatic leaves,while higher bacterial diversity was found in symptomatic leaves.After application,the diversity and abundance of microbial community structures in both types of leaves changed over time.Fungal microbiome communities showed greater sensitivity than bacterial groups,with the microbiome communities of asymptomatic leaves being more affected than those of symptomatic leaves.Fungal community diversity decreased for both symptomatic and asymptomatic leaves after 5 days of application,while the diversity of fungal community in symptomatic leaves showed an upward trend after 10 days of application.Meanwhile,bacterial community diversity increased in both symptomatic and asymptomatic leaves after 5 days of application but then declined in asymptomatic leaves after 15 days.The abundance of the dominant function group of phyllosphere bacteria(metabolism,genetic information processing,environmental information processing)was not affected by the application of Jiangong.However,the abundance of the dominant function group of phyllosphere fungi(animal pathogen-endophyte-wood saprotroph,endophyte-plant pathogen,plant pathogen-undefined saprotroph)was significantly affected by the application of Jiangong,and high variation was found in symptomatic leaves than that of asymptomatic leaves.The application of Jiangong-induced alterations in the community structure of the tobacco phyllosphere microbiome provides a basis for future tobacco brown spot control strategies based on phyllospheric microecology.
文摘Objective In kinesin-3,the neck coil correlates with the following segments to form an extended neck that contains a characteristic hinge diverse from a proline in KIF13B to a long flexible linker in KIF1A.The function of this neck hinge for controlling processive movement,however,remains unclear.Methods We made a series of modifications to the neck hinges of KIF13B and KIF1A and tested their movement using a single-molecule motility assay.Results In KIF13B,the insertion of flexible residues before or after the proline differentially impacts the processivity or velocity,while the removal of this proline increases the both.In KIF1A,the deletion of entire flexible neck hinge merely enhances the processivity.The engineering of these hinge-truncated necks of kinesin-3 into kinesin-1 similarly boosts the processive movement of kinesin-1.Conclusion The neck hinge in kinesin-3 controls its processive movement and proper modifications tune the motor motility,which provides a novel strategy to reshape the processive movement of kinesin motors.
文摘Objective Detection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability,health,and metabolic rate.After exposure to environmental stimuli,both the internal reference gene and target gene would be degraded.As a result,it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation.This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment.Methods The new RNA was labeled with 5-ethyluridine(5-EU)instead of uracil,and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of“Click Chemistry”and magnetic bead screening.Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon(M.B.)and quantitative reverse transcription PCR(qRT-PCR).Results The bacterial nascent RNA captured by“Click Chemistry”screening can be used as a reverse transcription template to form cDNA.Combined with the fluorescent molecular beacon M.B.1,the synthesis rate of rRNA at 37℃is 1.2 times higher than that at 15℃.The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃and 16℃when analyzed with nascent RNA rather than total RNA,enabling accurate detection of RNA transcription rates.Conclusion Compared to other article reported experimental methods that utilize screening magnetic columns,the technical scheme employed in this study is more suitable for bacteria,and the operation steps are simple and easy to implement,making it an effective RNA capture method for researchers.
基金Supported by the Natural Science Foundation of Heilongjiang Province(LH2021C032)。
文摘Avian influenza virus is one of the main pathogens causing avian influenza.In this experiment,the avian influenza hemagglutinin(HA)gene was transferred into tobacco by Agrobacterium tumefaciens-mediated method using kanamycin as a resistance marker to produce HA type edible vaccine designed for avian influenza hemagglutinin protein.The fusion of cholera toxin B subgene(CTB)and avian influenza HA was initiated by CaMV35S promoter,and then transformed into Nicotiana benthamiana to induce callus and adventitious bud differentiation,bud elongation,and growth and root induction.The total genomic DNA of 28 transgenic tobacco plants was extracted,and the CTB sequence of the CTB-HA fusion gene was used as a template to design primers for PCR amplification.Eight of them were positive,and four of them were expressed at the RNA level after RNA extraction and RT-PCR amplification.In western blot detection of protein extraction,two strains were shown at the position of the target protein.The results provided material support for the preparation of a transgenic plant oral vaccine against HA infection,and provided a theoretical basis for accelerating the development of a transgenic plant vaccine.
基金Supported by the Breeding Techniques for New Varieties of Acanthopanax senticosus(CZKYF2022-1-B023)。
文摘In order to optimize the ultrasonic extraction technique for the total flavonoid of leaf yellows plus, the contents of 21 leaf yellows plus total flavonoid from four regions in Heilongjiang Province were comparatively analyzed. The ultrasonic extraction technology was optimized by Box-Behnken response surface method, and the total flavonoid content of 21 kinds of Acanthopanax senticosus(Rupr. et Maxim.) Harms from different producing areas were analyzed by cluster analysis. The optimal process conditions were determined as ultrasonic time 30 min, solid-liquid ratio 1 : 12 and ultrasonic power 250 W, and the average yield of the total flavonoid was 1.453 mg·g^ (-1). By optimizing the ultrasonic-assisted extraction method, the total flavonoid content from different producing areas was compared in the experiment, which provided certain data support for the optimization of the extraction process in the future and laid a certain theoretical foundation for the quality analysis of Chinese medicinal materials.
基金funded by National Natural Science Foundation of China(No.32102214)Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences+1 种基金China Agriculture Research System(CARS-15-21)National Key R&D Program of China(2022YFD1400300)。
文摘Background Aphis gossypii(Hemiptera:Aphididae)is a worldwide polyphagous phloem-feeding agricultural pest,and it can produce offspring by sexual or asexual reproduction.Compared with dozens of generations by parthenogenesis,sexual reproduction is performed in only one generation within one year,and little is known about the sexual reproduction of A.gossypii.In this study,sexual females of A.gossypii were successfully obtained through a previously established induction platform,and the morphological characteristics,developmental dynamics,and temporal gene expression were examined.Subsequently,signaling pathways potentially involved in regulating the growth,development,and reproduction of sexual females were investigated.Results The morphological observation showed that from the 1st instar nymph to adult,sexual females exhibited a gradually deepened body color,an enlarged body size,longer antennae with a blackened end,and obviously protruding cauda(in adulthood).The anatomy found that the ovaries of sexual females developed rapidly from the 2^(nd)instar nymph,and the embedded oocytes matured in adulthood.In addition,time-course transcriptome analysis revealed that gene expression profiles across the development of sexual females fell into 9 clusters with distinct patterns,in which gene expression levels in clusters 1,5,and 8 peaked at the 2^(nd)instar nymphal stage with the largest number of up-regulated genes,suggesting that the 2^(nd)instar nymph was an important ovary development period.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis revealed that a large number of genes in the sexual female adult were enriched in the TGF-beta signaling pathway and Forkhead box O(FoxO)signaling pathway,highlighting their important role in sexual female adult development and reproduction.Conclusion The morphological changes of the sexual female at each developmental stage were revealed for the first time.In addition,time-course transcriptomic analyses suggest genes enriched in the TGF-beta signaling pathway and FoxO signaling pathway probably contribute to regulating the development and oocyte maturation of sexual females.Overall,these findings will facilitate the regulating mechanism research in the growth and development of sexual females by providing candidate genes.
文摘A novel 3D metal-organic framework(MOF)[Pr_(2)(L)_(3)(H_(2)O)5·H_(2)O]n(Pr-1),(H_(2)L=4,4'-oxybis(benzoic acid))with a rare structure of broken layer net,was constructed under the condition of solvothermal synthesis.The struc-ture and crystal net were analyzed and characterized.This rod net of Pr-1 is new to both RCSR and ToposPro data-bases,and is named as rn-12 as suggested.Due to the luminescent properties of H_(2)L and Pr(Ⅲ),the solid-state fluo-rescence property and sensing performance(solvents and metal ions)of Pr-1 were investigated.The sensing experi-ments indicated that Pr-1 could act as a fluorescence sensor to detect Cd^(2+)ions with good sensitivity.In addition,antibacterial activities show that Pr-1 exhibited stronger antibacterial activity against Escherichia coli(E.coli),Staphylococcus aureus(S.aureus),and Bacillus subtilis(B.subtilis)compared to synthetic materials.
文摘Objective Hepatocyte nuclear factor 4-alpha(HNF4A)is a critical transcription factor in the liver and pancreas.Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1(MODY1).Notably,MODY1 patients with HNF4A pathogenic mutations exhibit decreased responses to arginine and reduced plasma triglyceride levels,but the mechanisms remain unclear.This study aims to investigate the potential target genes transcriptionally regulated by HNF4A and explore its role in these metabolic pathways.Methods A stable 293T cell line expressing the HNF1A reporter was overexpressed with HNF4A.RNA sequencing(RNA-seq)was performed to analyze transcriptional differences.Transcription factor binding site prediction was then conducted to identify HNF4A binding motifs in the promoter regions of relevant target genes.Results RNA-seq results revealed a significant upregulation of transmembrane 4 L six family member 5(TM4SF5)mRNA in HNF4A-overexpressing cells.Transcription factor binding predictions suggested the presence of five potential HNF4A binding motifs in the TM4SF5 promoter.Finally,we confirmed that the DR1 site in the-57 to-48 region of the TM4SF5 promoter is the key binding motif for HNF4A.Conclusion This study identified TM4SF5 as a target gene of HNF4A and determined the key binding motif involved in its regulation.Given the role of TM4SF5 as an arginine sensor in mTOR signaling activation and triglyceride secretion,which closely aligns with phenotypes observed in MODY1 patients,our findings provide novel insights into the possible mechanisms by which HNF4A regulates triglyceride secretion in the liver and arginine-stimulated insulin secretion in the pancreas.
文摘Objective The nucleolar protein PES1(Pescadillo homolog 1)plays critical roles in ribosome biogenesis and cell cycle regulation,yet its involvement in cellular senescence remains poorly understood.This study aimed to comprehensively investigate the functional consequences of PES1 suppression in cellular senescence and elucidate the molecular mechanisms underlying its regulatory role.Methods Initially,we assessed PES1 expression patterns in two distinct senescence models:replicative senescent mouse embryonic fibroblasts(MEFs)and doxorubicin-induced senescent human hepatocellular carcinoma HepG2 cells.Subsequently,PES1 expression was specifically downregulated using siRNA-mediated knockdown in these cell lines as well as additional relevant cell types.Cellular proliferation and senescence were assessed by EdU incorporation and SA-β-gal staining assays,respectively.The expression of senescence-associated proteins(p53,p21,and Rb)and SASP factors(IL-6,IL-1β,and IL-8)were analyzed by Western blot or qPCR.Furthermore,Northern blot and immunofluorescence were employed to evaluate pre-rRNA processing and nucleolar morphology.Results PES1 expression was significantly downregulated in senescent MEFs and HepG2 cells.PES1 knockdown resulted in decreased EdU-positive cells and increased SA-β-gal-positive cells,indicating proliferation inhibition and senescence induction.Mechanistically,PES1 suppression activated the p53-p21 pathway without affecting Rb expression,while upregulating IL-6,IL-1β,and IL-8 production.Notably,PES1 depletion impaired pre-rRNA maturation and induced nucleolar stress,as evidenced by aberrant nucleolar morphology.Conclusion Our findings demonstrate that PES1 deficiency triggers nucleolar stress and promotes p53-dependent(but Rb-independent)cellular senescence,highlighting its crucial role in maintaining nucleolar homeostasis and regulating senescence-associated pathways.
基金Supported by the Academic Backbone Fund of Northeast Agricultural University(19XG20)the Excellent Young Scholars Fund of Harbin Medical University(HYD2020JQ0016)。
文摘Esophageal cancer(EC)is one of the most common malignancies in the world,and there is no specific treatment drug for esophageal cancer yet.Doramectin(DRM)is a broad-spectrum anti-parasitic drug,and it plays an important role in the treatment of animal diseases,while DRM has not been reported for the treatment of esophageal squamous cell carcinoma(ESCC).The purpose of this study was to investigate the anticancer effects and potential molecular mechanisms of DRM in ESCC.In the present study,the impact of DRM on the viability of ESCC was examined by methylthiazolyldiphenyl-tetrazolium bromide(MTT).Autophagy was measured by transmission electron microscopy(TEM),Western blot and immunohistochemistry.The apoptosis rate was measured by Western blot,flow cytometry and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL).Meanwhile,autophagy inhibition was achieved by using chloroquine(CQ).After autophagy inhibition,cell proliferation and cloning ability were significantly inhibited,and the expression level of apoptotic protein was significantly changed compared with that of DRM alone.Additionally,Eca109-derived xenografts were established for testing the DRM-induced autophagy in vivo.It was found that DRM significantly inhibited the proliferation of Eca109 and EC9706 cells in vitro and in vivo in a dose-dependent manner by activating autophagy.DRM was able to significantly repress colony formation in Eca109 and EC9706 cells in vitro.At the same time,DRM could induce apoptosis of ESCC in vitro,it was also regulated through mitochondrial pathways.Meanwhile,DRM induced autophagy and inhibited the proliferation of ESCC,and exhibited little toxicity in organs in vivo.Moreover,DRM-induced autophagy could inhibit the apoptosis of EC in vitro and in vivo.Further experiment suggested that DRM might induce autophagy by the Akt/mTOR pathway.In conclusion,the present study was the first to clarify that DRM could inhibit Eca109 and EC9706 cells proliferation through activating autophagy by the Akt/mTOR pathway.DRM might be a potentially effective treatment for EC.
文摘Platycodon grandiflorum A.DC.(PAl)C)root was taken as experiment material to extract polysaccharide.On the base of single factor tests(extraction time,extraction temperature,liquid-solid ratio,solvent pH value and NaCl concentration),the study concluded the main factors affecting the extraction of PADC polysaccharide,which are liquid-solid ratio,extraction time and extraction temperature.Then through central-composite test design,the extraction conditions were concluded as liquid-solid ratio 34.43,extraction time 89.83 min and extraction temperature 52.47℃.By means of validation experiments,the adequacy of this model was confirmed.
基金Project(200501) supported the "985" Program of China
文摘Starch-nanoparticles were synthesized in water-in-oil microemusion at room temperature, and the starch-nanoparticles were coated with poly-L-lysine. The surface of the starch-nanoparticles was combined with fluorescence material Ru(bpy)32+·6H2O, and then the particles were characterized via transmission electron microscope. The fluorescence nanoparticles were conjugated with plasmid DNA to form complexes, and then treated with ultrasound and DNase I. pEGAD plasmid DNA-nanoparticle complexes were co-cultured with plant suspension cells of Dioscrea Zigiberensis G H Wright, and treated with ultrasound. The results show that the diameter of the fluorescence starch-nanoparticles is 50-100 nm. DNA-nanoparticle complexes can protect DNA from ultrasound damage as well as from DNase I cleavage. Mediated by ultrasound, pEGAD plasmid DNA-nanoparticle complexes can pierce into the cell wall, cell membrane and nucleus membrane of plant suspension cells. The green fluorescence protein(GFP) gene at a high frequency exceeds 5%. This nano-biomaterial can efficiently solve the problem that exterior genes cannot traverse the plant cell wall easily.
基金Supported by"863"Project of Ministry of Science and Technology of China(2013AA102504-03)
文摘Methionine （Met） and lysine （Lys） have been reported as the first two limiting amino acids （AA） for maximum milk yield and milk protein production. Supplying these AA may improve microbial protein synthesis and therefore improve milk production without adding excess N to the environment. This observation utilized fermented soybean meal （SBM）, cottonseed meal （CSM）, rapeseed meal （RSM） and corn by Bacillus subtilis 168 and Leuconostoc mesenteroides as core feedstuffs to produce special biological protein feed for dairy cow. The results showed that the milk production, milk protein percentage, milk fat percentage and milk DM percentage of test groups in trial period were significantly more than those of the control group （P〈0.01）, the results showed that adding fermenting protein feed in dairy cow diets could significantly improve milk yield, milk protein and milk fat content. The economic benefits of actual application were analyzed, the group of 0.5% was the best compared with the other groups.
