This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogeno...This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogenous substances to efficient cultivationof Isatis indigotica. Polyethylene glycol (PEG) was used to simulate drought stress to deal with seeds of Isatis indigotica at thegermination stage (concentration: 0, 10%, 15%, and 20%). Simultaneous operation of exogenous growth regulators [microbialinoculum (MI), γ-aminobutyric acid (GABA) and salicylic acid (SA)] and PEG were implemented in seeds of Isatis indigotica.The effects of drought stress and the mitigation of exogenous substances were observed by statistics of seed germination potential,germination rate, hypocotyl length, and radicle length of each treatment. The effects of exogenous substances on the content ofalkaloids, crude protein and free amino acids in the leaves of Isatis indigotica grown in a greenhouse were determined after sprayingexogenous substances on the plants. The differences of germination potential, germination rate, hypocotyl length, and radicle lengthamong 15% PEG stress treatment, 10% PEG stress treatment and the control were significant (P<0.05). According to the predesignedgermination standard, the seeds did not germinate under 20% PEG stress treatment. When the PEG concentration was 15%, the resultsof seed germination potential and germination rate after adding MI were significantly different from those under stress alone (P<0.05).When exposed to 10% PEG stress, the supplementation of GABA led to a notable increase in radicle length of Isatis indigotica seeds,showing significant differences compared to other three treatments. The application of MI and GABA under 15% PEG stress resultedin a significant increase in the radicle and hypocotyl length of Isatis indigotica seeds compared to other two treatments. The contentof the total alkaloids in leaves of Isatis indigotica was significantly increased after spraying GABA. Meanwhile, the contents of crudeprotein and the total free amino acids were kept constant after spraying exogenous substances. Application of MI and GABA couldalleviate drought stress of Isatis indigotica. The content of the total alkaloids in leaves of Isatis indigotica could significantly increaseafter spraying GABA.展开更多
The fresh leaves and oven dried leaves from three varieties of black currant (Brodtrop,liangye,Risager)were analysed for the total contents of flavonoids by spectrophotometric methods in terms of rutin equivalent.In ...The fresh leaves and oven dried leaves from three varieties of black currant (Brodtrop,liangye,Risager)were analysed for the total contents of flavonoids by spectrophotometric methods in terms of rutin equivalent.In addition,antioxidant activities of different addition amounts of extracts of black currant (Brodtrop)leaves in lard were measured by the Oven-Storage Test,and compared with the synthetic antioxidants.The results showed the contents of the flavonoids were very rich in the leaves of three varieties from black currant.Among them the leaves of Brodtrop had the highest contents of flavonoids followed by liangye and Risager.The total contents of flavonoids in fresh leaves reach 231 70 mg/100 g,170 01 mg/100 g and 78 07 mg/100 g respectively.In the same circumstance the oven dried leaves were 397 82 mg/100 g,275 89 mg/100 g and 169 73 mg/100 g.The extracts of the leaves from Brodtrop exhibited strong antioxidant activities on lard,which increased with the addition amount of the flavonoid extracts.The antioxidant activities decreased in the order of 0 02% quercetin >0 02% BHT(butylated hydroxy toluene)>0 40% extracts>0 20% extracts>0 10% extracts>0 05% extracts>0 02% extracts>control.展开更多
Employing the orthogonal design,the optimal extraction conditions for flavonoid in leaves of black currant were determined by colorimetric estimation,which were extraction agent of 75% EtOH-H 2O,the material ratio(ra...Employing the orthogonal design,the optimal extraction conditions for flavonoid in leaves of black currant were determined by colorimetric estimation,which were extraction agent of 75% EtOH-H 2O,the material ratio(ratio of weight of fresh leaves to volume of the solvent)of 1:16,the temperature of 35℃,the time of 1 h and times of three.A total extraction rate of over 97% and the flavonoid contents of 179 33 mg/100 g were obtained under the optimal extraction conditions.展开更多
The aims were to investigate the effect of salt stress on key enzyme activity of nitrogen metabolism and the concentration of nitrate nitrogen and ammonium nitrogen response to salt stress.Two rice cultivars,Mudanjian...The aims were to investigate the effect of salt stress on key enzyme activity of nitrogen metabolism and the concentration of nitrate nitrogen and ammonium nitrogen response to salt stress.Two rice cultivars,Mudanjiang 30(sensitive cultivar)and Longdao 5(salt-tolerant cultivar),were treated with different salt concentrations(CK 0%,S10.075%,S20.15%,S30.225%and S40.3%).The results showed that the activities of nitrate reductase(NR),glutamine synthase(GS),glutamate synthase(GOGAT)and glutamate dehydrogenase(GDH)in the functional leaves and roots of rice in cold region presented a single peak curve change and the peak occurred in the heading stage;compared with those of the CK,the activities of NR,GS and GOGAT of rice in cold region decreased,but the activity of GDH increased in the heading stage under salt stress.The variation for key enzyme activity of nitrogen metabolism was the highest under S4 treatment.The activities of NR,GS and GOGAT in the functional leaves significantly decreased compared with those in roots;the concentrations of nitrate nitrogen and ammonium nitrogen in the functional leaves and roots of rice in cold region presented a single peak curve change and the peak occurred in the heading stage;compared with that of the CK,the concentration of nitrate nitrogen decreased in leaves and roots,the concentration of ammonium nitrogen decreased and the concentration of ammonium nitrogen in roots increased under salt stress.The variations for the activities of NR,GS and GOGAT in the functional leaves and roots of Longdao 5 were less than those of Mudanjiang 30 under the same concentration of salt stress.展开更多
People consume tea brewed from the leaves of the Camellia sinensis plant for about 50 centuries. Health benefits of the tea have been investigated for about three decades. Especially green tea shows antitoxic and lots...People consume tea brewed from the leaves of the Camellia sinensis plant for about 50 centuries. Health benefits of the tea have been investigated for about three decades. Especially green tea shows antitoxic and lots of properties with its determined ingredients. Turkey is not only the one of the best consumer and but also good producer of the tea as being 5 th producer all over the world. It grows eastern region of the Turkey and high quality tea is imported and exported. To have quality tea, grooving soils are also crucial. In the current research, Tea leaves and their own grown soils were collected from 20 stations where the most tea producer cities as Trabzon, Rize and Artvin tea fields of the eastern of Black Sea Region in Turkey. The cultivated tea and their own grown soil samples were analysed by using EDXRF Spectrometry. In the soil samples, the elements Mg, Al, Si, P, S, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Sr, Ba, Pb on percent level and the elements Mg, Al, Si, P, Cl, K, Ca, Ti, Mn, Fe, Ni, Cu, Zn and Sr in the tea leaves were detected by using SKRAY 3600-EDXRF.and also the obtained data were evaluated with the Kriging interpolation of geostatistical method. Element content were investigated in the soil and tea samples according to the geological situations and also the relation of elemental difference between the tea and the own grown soil. Pb/Zn ratio was also anaysed in the samples.展开更多
A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/...A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/16-h dark) and long-day(16-h light/8-h dark) conditions.A total of 148 clones were sequenced,representing 76 unique ESTs which corresponded to about 20% of 738 clones from the cDNA library and showed a significant up-regulation of at least three fold verified by dot blot hybridization.The putative functions of ESTs were predicted by Blastn and Blastx.The 43 differentially expressed genes identified by subtractions were classified according to their putative functions generated by Blast analysis.Genetic functional analysis indicated that putative proteins encoded by these genes were related to diverse functions during organism development,which include biological regulation pathways such as transcription,signal transduction and programmed cell death,protein,nucleic acid and carbohydrate macromolecule degradation,the cell wall modification,primary and secondary metabolism and stress response.Two soybean transcription factors enhanced in SD conditions,GAMYB-binding protein and DNA binding protein RAV cDNAs that may be involved in SD soybean photoperiod response,had been isolated using 5'-and 3'-rapid amplification of cDNA ends(RACE)(Genbank Accession numbers DQ112540 and DQ147914).展开更多
Objective:Middle ear cholesteatoma is a non-tumorous condition that typically leads to hearing loss,bone destruction,and other severe complications.Despite surgery being the primary treatment,the recurrence rate remai...Objective:Middle ear cholesteatoma is a non-tumorous condition that typically leads to hearing loss,bone destruction,and other severe complications.Despite surgery being the primary treatment,the recurrence rate remains high.Therefore,exploring the molecular mechanisms underlying cholesteatoma is crucial for discovering new therapeutic approaches.This study aims to explore the involvement of N6-methyladenosine(m^(6)A)methylation in long non-coding RNAs(lncRNAs)in the biological functions and related pathways of middle ear cholesteatoma.Methods:The m^(6)A modification patterns of lncRNA in middle ear cholesteatoma tissues(n=5)and normal post-auricular skin tissues(n=5)were analyzed using an lncRNA m^(6)A transcriptome microarray.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses were conducted to identify potential biological functions and signaling pathways involved in the pathogenesis of middle ear cholesteatoma.Methylated RNA immunoprecipitation(MeRIP)-PCR was used to validate the m^(6)A modifications in cholesteatoma and normal skin tissues.Results:Compared with normal skin tissues,1525 lncRNAs were differentially methylated in middle ear cholesteatoma tissues,with 1048 showing hypermethylation and 477 showing hypomethylation[fold change(FC)≥3 or<1/3,P<0.05].GO enrichment analysis indicated that hypermethylated lncRNAs were involved in protein phosphatase inhibitor activity,neuron-neuron synapse,and regulation ofα-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid(AMPA)receptor activity.Hypomethylated lncRNAs were associated with mRNA methyltransferase activity,secretory granule membrane,and mRNA methylation.KEGG analysis revealed that hypermethylated lncRNAs were mainly associated with 5 pathways:the Hedgehog signaling pathway,viral protein interaction with cytokines and cytokine receptors,mitogen-activated protein kinase(MAPK)signaling pathway,cytokine-cytokine receptor interaction,and adrenergic signaling in cardiomyocytes.Hypomethylated lncRNAs were mainly involved in 4 pathways:Renal cell carcinoma,tumor necrosis factor signaling pathway,transcriptional misregulation in cancer,and cytokine-cytokine receptor interaction.Additionally,MeRIP-PCR confirmed the changes in m^(6)A methylation levels in NR_033339,NR_122111,NR_130744,and NR_026800,consistent with microarray analysis.Real-time PCR also confirmed the significant upregulation of MAPK1 and NF-κB,key genes in the MAPK signaling pathway.Conclusion:This study reveals the m^(6)A modification patterns of lncRNAs in middle ear cholesteatoma,suggests a direction for further research into the role of lncRNA m^(6)A modification in the etiology of cholesteatoma.The findings provide potential therapeutic targets for the treatment of middle ear cholesteatoma.展开更多
基金the Doctoral Research Initiation Foundation of Changzhi Medical College(BS202005)。
文摘This study was to investigate the effects of three exogenous substances on chemical constituents of Isatis indigotica leavesand their efficacy in alleviating drought stress, and explore the methods of applying exogenous substances to efficient cultivationof Isatis indigotica. Polyethylene glycol (PEG) was used to simulate drought stress to deal with seeds of Isatis indigotica at thegermination stage (concentration: 0, 10%, 15%, and 20%). Simultaneous operation of exogenous growth regulators [microbialinoculum (MI), γ-aminobutyric acid (GABA) and salicylic acid (SA)] and PEG were implemented in seeds of Isatis indigotica.The effects of drought stress and the mitigation of exogenous substances were observed by statistics of seed germination potential,germination rate, hypocotyl length, and radicle length of each treatment. The effects of exogenous substances on the content ofalkaloids, crude protein and free amino acids in the leaves of Isatis indigotica grown in a greenhouse were determined after sprayingexogenous substances on the plants. The differences of germination potential, germination rate, hypocotyl length, and radicle lengthamong 15% PEG stress treatment, 10% PEG stress treatment and the control were significant (P<0.05). According to the predesignedgermination standard, the seeds did not germinate under 20% PEG stress treatment. When the PEG concentration was 15%, the resultsof seed germination potential and germination rate after adding MI were significantly different from those under stress alone (P<0.05).When exposed to 10% PEG stress, the supplementation of GABA led to a notable increase in radicle length of Isatis indigotica seeds,showing significant differences compared to other three treatments. The application of MI and GABA under 15% PEG stress resultedin a significant increase in the radicle and hypocotyl length of Isatis indigotica seeds compared to other two treatments. The contentof the total alkaloids in leaves of Isatis indigotica was significantly increased after spraying GABA. Meanwhile, the contents of crudeprotein and the total free amino acids were kept constant after spraying exogenous substances. Application of MI and GABA couldalleviate drought stress of Isatis indigotica. The content of the total alkaloids in leaves of Isatis indigotica could significantly increaseafter spraying GABA.
文摘The fresh leaves and oven dried leaves from three varieties of black currant (Brodtrop,liangye,Risager)were analysed for the total contents of flavonoids by spectrophotometric methods in terms of rutin equivalent.In addition,antioxidant activities of different addition amounts of extracts of black currant (Brodtrop)leaves in lard were measured by the Oven-Storage Test,and compared with the synthetic antioxidants.The results showed the contents of the flavonoids were very rich in the leaves of three varieties from black currant.Among them the leaves of Brodtrop had the highest contents of flavonoids followed by liangye and Risager.The total contents of flavonoids in fresh leaves reach 231 70 mg/100 g,170 01 mg/100 g and 78 07 mg/100 g respectively.In the same circumstance the oven dried leaves were 397 82 mg/100 g,275 89 mg/100 g and 169 73 mg/100 g.The extracts of the leaves from Brodtrop exhibited strong antioxidant activities on lard,which increased with the addition amount of the flavonoid extracts.The antioxidant activities decreased in the order of 0 02% quercetin >0 02% BHT(butylated hydroxy toluene)>0 40% extracts>0 20% extracts>0 10% extracts>0 05% extracts>0 02% extracts>control.
文摘Employing the orthogonal design,the optimal extraction conditions for flavonoid in leaves of black currant were determined by colorimetric estimation,which were extraction agent of 75% EtOH-H 2O,the material ratio(ratio of weight of fresh leaves to volume of the solvent)of 1:16,the temperature of 35℃,the time of 1 h and times of three.A total extraction rate of over 97% and the flavonoid contents of 179 33 mg/100 g were obtained under the optimal extraction conditions.
基金Supported by the National Key R&D Program of China(2016YFD0300104)。
文摘The aims were to investigate the effect of salt stress on key enzyme activity of nitrogen metabolism and the concentration of nitrate nitrogen and ammonium nitrogen response to salt stress.Two rice cultivars,Mudanjiang 30(sensitive cultivar)and Longdao 5(salt-tolerant cultivar),were treated with different salt concentrations(CK 0%,S10.075%,S20.15%,S30.225%and S40.3%).The results showed that the activities of nitrate reductase(NR),glutamine synthase(GS),glutamate synthase(GOGAT)and glutamate dehydrogenase(GDH)in the functional leaves and roots of rice in cold region presented a single peak curve change and the peak occurred in the heading stage;compared with those of the CK,the activities of NR,GS and GOGAT of rice in cold region decreased,but the activity of GDH increased in the heading stage under salt stress.The variation for key enzyme activity of nitrogen metabolism was the highest under S4 treatment.The activities of NR,GS and GOGAT in the functional leaves significantly decreased compared with those in roots;the concentrations of nitrate nitrogen and ammonium nitrogen in the functional leaves and roots of rice in cold region presented a single peak curve change and the peak occurred in the heading stage;compared with that of the CK,the concentration of nitrate nitrogen decreased in leaves and roots,the concentration of ammonium nitrogen decreased and the concentration of ammonium nitrogen in roots increased under salt stress.The variations for the activities of NR,GS and GOGAT in the functional leaves and roots of Longdao 5 were less than those of Mudanjiang 30 under the same concentration of salt stress.
文摘People consume tea brewed from the leaves of the Camellia sinensis plant for about 50 centuries. Health benefits of the tea have been investigated for about three decades. Especially green tea shows antitoxic and lots of properties with its determined ingredients. Turkey is not only the one of the best consumer and but also good producer of the tea as being 5 th producer all over the world. It grows eastern region of the Turkey and high quality tea is imported and exported. To have quality tea, grooving soils are also crucial. In the current research, Tea leaves and their own grown soils were collected from 20 stations where the most tea producer cities as Trabzon, Rize and Artvin tea fields of the eastern of Black Sea Region in Turkey. The cultivated tea and their own grown soil samples were analysed by using EDXRF Spectrometry. In the soil samples, the elements Mg, Al, Si, P, S, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Sr, Ba, Pb on percent level and the elements Mg, Al, Si, P, Cl, K, Ca, Ti, Mn, Fe, Ni, Cu, Zn and Sr in the tea leaves were detected by using SKRAY 3600-EDXRF.and also the obtained data were evaluated with the Kriging interpolation of geostatistical method. Element content were investigated in the soil and tea samples according to the geological situations and also the relation of elemental difference between the tea and the own grown soil. Pb/Zn ratio was also anaysed in the samples.
文摘A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/16-h dark) and long-day(16-h light/8-h dark) conditions.A total of 148 clones were sequenced,representing 76 unique ESTs which corresponded to about 20% of 738 clones from the cDNA library and showed a significant up-regulation of at least three fold verified by dot blot hybridization.The putative functions of ESTs were predicted by Blastn and Blastx.The 43 differentially expressed genes identified by subtractions were classified according to their putative functions generated by Blast analysis.Genetic functional analysis indicated that putative proteins encoded by these genes were related to diverse functions during organism development,which include biological regulation pathways such as transcription,signal transduction and programmed cell death,protein,nucleic acid and carbohydrate macromolecule degradation,the cell wall modification,primary and secondary metabolism and stress response.Two soybean transcription factors enhanced in SD conditions,GAMYB-binding protein and DNA binding protein RAV cDNAs that may be involved in SD soybean photoperiod response,had been isolated using 5'-and 3'-rapid amplification of cDNA ends(RACE)(Genbank Accession numbers DQ112540 and DQ147914).
基金supported by the National Natural Science Foundation(82071036,82000973)the Natural Science Foundation of Hunan Province(2022JJ30821,2019JJ50967)the Special Project for the Construction of Innovative Provinces in Hunan Province(2023SK4030),China。
文摘Objective:Middle ear cholesteatoma is a non-tumorous condition that typically leads to hearing loss,bone destruction,and other severe complications.Despite surgery being the primary treatment,the recurrence rate remains high.Therefore,exploring the molecular mechanisms underlying cholesteatoma is crucial for discovering new therapeutic approaches.This study aims to explore the involvement of N6-methyladenosine(m^(6)A)methylation in long non-coding RNAs(lncRNAs)in the biological functions and related pathways of middle ear cholesteatoma.Methods:The m^(6)A modification patterns of lncRNA in middle ear cholesteatoma tissues(n=5)and normal post-auricular skin tissues(n=5)were analyzed using an lncRNA m^(6)A transcriptome microarray.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses were conducted to identify potential biological functions and signaling pathways involved in the pathogenesis of middle ear cholesteatoma.Methylated RNA immunoprecipitation(MeRIP)-PCR was used to validate the m^(6)A modifications in cholesteatoma and normal skin tissues.Results:Compared with normal skin tissues,1525 lncRNAs were differentially methylated in middle ear cholesteatoma tissues,with 1048 showing hypermethylation and 477 showing hypomethylation[fold change(FC)≥3 or<1/3,P<0.05].GO enrichment analysis indicated that hypermethylated lncRNAs were involved in protein phosphatase inhibitor activity,neuron-neuron synapse,and regulation ofα-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid(AMPA)receptor activity.Hypomethylated lncRNAs were associated with mRNA methyltransferase activity,secretory granule membrane,and mRNA methylation.KEGG analysis revealed that hypermethylated lncRNAs were mainly associated with 5 pathways:the Hedgehog signaling pathway,viral protein interaction with cytokines and cytokine receptors,mitogen-activated protein kinase(MAPK)signaling pathway,cytokine-cytokine receptor interaction,and adrenergic signaling in cardiomyocytes.Hypomethylated lncRNAs were mainly involved in 4 pathways:Renal cell carcinoma,tumor necrosis factor signaling pathway,transcriptional misregulation in cancer,and cytokine-cytokine receptor interaction.Additionally,MeRIP-PCR confirmed the changes in m^(6)A methylation levels in NR_033339,NR_122111,NR_130744,and NR_026800,consistent with microarray analysis.Real-time PCR also confirmed the significant upregulation of MAPK1 and NF-κB,key genes in the MAPK signaling pathway.Conclusion:This study reveals the m^(6)A modification patterns of lncRNAs in middle ear cholesteatoma,suggests a direction for further research into the role of lncRNA m^(6)A modification in the etiology of cholesteatoma.The findings provide potential therapeutic targets for the treatment of middle ear cholesteatoma.
