Background:The Emei Shan Liocichla(Liocichla omeiensis) is an endemic bird species to southwestern China with a small geographic range. However, little was known about the genetic status of this threatened species.Met...Background:The Emei Shan Liocichla(Liocichla omeiensis) is an endemic bird species to southwestern China with a small geographic range. However, little was known about the genetic status of this threatened species.Methods:We applied restriction-site-associated DNA sequencing(RAD-Seq) for rapid mass identification of microsatellite markers of the Emei Shan Liocichla.Results:A total of 11,564 microsatellite sequences were obtained, 600 random loci were designed for screening and 24 polymorphic microsatellite loci were selected for further validation. The average allele number, average observed heterozygosity and average expected heterozygosity were relatively low in our samples, which were 6.08, 0.6618 and 0.7048, respectively, indicating that the Emei Shan Liocichla might have lost some genetic diversity. Further analyses suggested that the populations distributed on two mountains(Daxiangling and Xiaoliangshan) showed a modest degree of genetic differentiation.Conclusions:These novel microsatellite markers provided valuable preliminary knowledge regarding the genetic status of the Emei Shan Liocichla and can be useful in further studies, as well as in the management and conservation of this species.展开更多
Deciphering the role of climatic oscillations in species divergence helps us understand the mechanisms that shape global biodiversity.The coldadapted species may have expanded their distribution with the development o...Deciphering the role of climatic oscillations in species divergence helps us understand the mechanisms that shape global biodiversity.The coldadapted species may have expanded their distribution with the development of glaciers during glacial period.With the retreat of glaciers,these species were discontinuously distributed in the high-altitude mountains and isolated by geographical barriers.However,the study that focuses on the speciation process of cold-adapted species is scant.To fill this gap,we combined population genetic data and ecological niche models(ENMs)to explore divergence process of snow partridge(Lerwa lerwa).Lerwa lerwa is a cold-adapted bird that is distributed from 4,000 to 5,500 m.We found 2 genetic populations within L.lerwa,and they diverged from each other at about 0.40–0.44 million years ago(inter-glacial period after Zhongliangan glaciation).The ENMs suggested that L.lerwa expanded to the low elevations of the Himalayas and Hengduan mountains during glacial period,whereas it contracted to the high elevations,southern of Himalayas,and Hengduan mountains during inter-glacial periods.Effective population size trajectory also suggested that L.lerwa expanded its population size during the glacial period.Consistent with our expectation,the results support that inter-glacial isolation contributed to the divergence of cold-adapted L.lerwa on Qinghai-Tibetan Plateau.This study deepens our understanding of how climatic oscillations have driven divergence process of cold-adapted Phasianidae species distributed on mountains.展开更多
Anorexia nervosa(AN)is a mental disorder,affecting mainly females in society.Understanding partially the factor of the disorder,researchers also designed several therapies to alleviate AN.Cognitivebehavioral therapy(C...Anorexia nervosa(AN)is a mental disorder,affecting mainly females in society.Understanding partially the factor of the disorder,researchers also designed several therapies to alleviate AN.Cognitivebehavioral therapy(CBT),which focuses on changing thought patterns of patients,and addressing inappropriate behaviors,is one of the beneficial therapies.In China,females whose disorder are caused by environmental factors are highly recommended to have CBT.To successfully realize the use of CBT to AN,we have to understand the strategy and treatment.展开更多
Objective:Hepatocellular carcinoma(HCC)is a leading cause of mortality worldwide.Huachansu(Cinobufacini)is active extract isolated from the dry skin of Bufo Bufo gargarizans.It has now been widely used in clinical tre...Objective:Hepatocellular carcinoma(HCC)is a leading cause of mortality worldwide.Huachansu(Cinobufacini)is active extract isolated from the dry skin of Bufo Bufo gargarizans.It has now been widely used in clinical treatment of cancer,this study is to clarify the material basis of down-regulation of thymidylate synthase(TYMS)induced by Huachansu.Methods:Our study utilized UPLC-MS/MS to identify major bioactive components from Huachansu.Cell Counting Kit 8(CCK-8)assay and clone formation assay were used to examine the cell viability of tumor cells.TYMS and y-H2AX level were detected by using quantitative real-time RT-PCR and/or western blotting.Small interfering RNA(siRNA)transfection was used to explore whether inhibition of TYMS could enhance the suppressive effect of Huachansu on cell growth of HCC cells.Results:In our study,firstly,we identify 21 major bioactive components from Huachansu.CCK-8 assay results showed that Huachansu and its bioactive bufadienolides(Bufalin,Bufotalin,Cinobufotalin,Desacetylcinobufagin,Arenobufagin,Telocinobu fagin,and Resibu fogenin)significantly inhibited the proliferation of HepG2 and SK-HEP-1 cells in a dose-and time-dependent manner.Further molecular mechanistic investigation demonstrates that Huachansu significantly suppresses thymidylate synthase(TYMS),the enzyme which provides the sole de novo source of thymidylate for DNA synthesis.The inhibition of TYMS could lead to cell-cycle block and DNA damage of HCC cells.Furthermore,we identified that Huachansu markedly increased y-H2AX expression,which indicated the presence of DNA damage.Moreover,we confirmed that transfection of cells with small interfering RNA specific to TYMS could increase the suppressive effects of Huachansu on the HCC cells proliferation.Quantitative RT-PCR analysis showed that Huachansu treatment had no effect on the transcription level of TYMS.Fu rthermore,proteasomal inhibitor MG132 could block TYMS inhibition induced by Huachansu,and concomita nt administration of protein synthesis inhibitor cycloheximide(CHX)with Huachansu could further suppress the protein level of TYMS,indicating that Huachansu promotes proteasome-dependent degradation of TYMS in liver cancer cells.More importantly,the bioactive bufadienolides of Huachansu such as Bufalin,Bufotalin,Cinobufotalin,Desacetylcinobufagin,Arenobufagin,Telocinobufagin,and Resibufogenin could also significantly restrain the protein level of TYMS,revealing the material basis of inhibition of TYMS exposed to Huachansu.5-Fluorouracil(5-FU)is a TYMS inhibitor,we also evaluate the effects of the combined treatment of Huachansu with 5-FU,the results show that interactions between Huachansu and 5-FU are synergistic or antagonistic.Thus,in clinical,attention should be paid to the dosage of Huachansu in combination with 5-FU.Conclusion:Huachansu inhibits the growth and induces DNA damage of human HCC cells through proteasome-dependent degradation of TYMS,bioactive bufadienolides are the material basis of downregulation of TYMS induced by Huachansu.展开更多
基金supported by the National Natural Science Foundation of China (Grant No. 31272330) to YFthe National Key Programme of Research and Development, Ministry of Science and Technology (2016YFC0503200) to ZZ
文摘Background:The Emei Shan Liocichla(Liocichla omeiensis) is an endemic bird species to southwestern China with a small geographic range. However, little was known about the genetic status of this threatened species.Methods:We applied restriction-site-associated DNA sequencing(RAD-Seq) for rapid mass identification of microsatellite markers of the Emei Shan Liocichla.Results:A total of 11,564 microsatellite sequences were obtained, 600 random loci were designed for screening and 24 polymorphic microsatellite loci were selected for further validation. The average allele number, average observed heterozygosity and average expected heterozygosity were relatively low in our samples, which were 6.08, 0.6618 and 0.7048, respectively, indicating that the Emei Shan Liocichla might have lost some genetic diversity. Further analyses suggested that the populations distributed on two mountains(Daxiangling and Xiaoliangshan) showed a modest degree of genetic differentiation.Conclusions:These novel microsatellite markers provided valuable preliminary knowledge regarding the genetic status of the Emei Shan Liocichla and can be useful in further studies, as well as in the management and conservation of this species.
基金supported by the second National Survey on Terrestrial Wildlife Resources in China.
文摘Deciphering the role of climatic oscillations in species divergence helps us understand the mechanisms that shape global biodiversity.The coldadapted species may have expanded their distribution with the development of glaciers during glacial period.With the retreat of glaciers,these species were discontinuously distributed in the high-altitude mountains and isolated by geographical barriers.However,the study that focuses on the speciation process of cold-adapted species is scant.To fill this gap,we combined population genetic data and ecological niche models(ENMs)to explore divergence process of snow partridge(Lerwa lerwa).Lerwa lerwa is a cold-adapted bird that is distributed from 4,000 to 5,500 m.We found 2 genetic populations within L.lerwa,and they diverged from each other at about 0.40–0.44 million years ago(inter-glacial period after Zhongliangan glaciation).The ENMs suggested that L.lerwa expanded to the low elevations of the Himalayas and Hengduan mountains during glacial period,whereas it contracted to the high elevations,southern of Himalayas,and Hengduan mountains during inter-glacial periods.Effective population size trajectory also suggested that L.lerwa expanded its population size during the glacial period.Consistent with our expectation,the results support that inter-glacial isolation contributed to the divergence of cold-adapted L.lerwa on Qinghai-Tibetan Plateau.This study deepens our understanding of how climatic oscillations have driven divergence process of cold-adapted Phasianidae species distributed on mountains.
文摘Anorexia nervosa(AN)is a mental disorder,affecting mainly females in society.Understanding partially the factor of the disorder,researchers also designed several therapies to alleviate AN.Cognitivebehavioral therapy(CBT),which focuses on changing thought patterns of patients,and addressing inappropriate behaviors,is one of the beneficial therapies.In China,females whose disorder are caused by environmental factors are highly recommended to have CBT.To successfully realize the use of CBT to AN,we have to understand the strategy and treatment.
基金financially supported by National Natural Science Foundation of China Grant(No.82204693)Taishan scholar engineering youth expert program of Shandong Province(No.tsqn202103110)+2 种基金Young and Creative Team for Talent Introduction of Shandong Province(No.10073004)Binzhou Medical University Scientific Research Fund for High-level Talents(NO.BY2020KYQD13,2019KYQD06)Standard study on Chinese medicinal materials and Decoction pieces in Shandong Province(No.2020-201)。
文摘Objective:Hepatocellular carcinoma(HCC)is a leading cause of mortality worldwide.Huachansu(Cinobufacini)is active extract isolated from the dry skin of Bufo Bufo gargarizans.It has now been widely used in clinical treatment of cancer,this study is to clarify the material basis of down-regulation of thymidylate synthase(TYMS)induced by Huachansu.Methods:Our study utilized UPLC-MS/MS to identify major bioactive components from Huachansu.Cell Counting Kit 8(CCK-8)assay and clone formation assay were used to examine the cell viability of tumor cells.TYMS and y-H2AX level were detected by using quantitative real-time RT-PCR and/or western blotting.Small interfering RNA(siRNA)transfection was used to explore whether inhibition of TYMS could enhance the suppressive effect of Huachansu on cell growth of HCC cells.Results:In our study,firstly,we identify 21 major bioactive components from Huachansu.CCK-8 assay results showed that Huachansu and its bioactive bufadienolides(Bufalin,Bufotalin,Cinobufotalin,Desacetylcinobufagin,Arenobufagin,Telocinobu fagin,and Resibu fogenin)significantly inhibited the proliferation of HepG2 and SK-HEP-1 cells in a dose-and time-dependent manner.Further molecular mechanistic investigation demonstrates that Huachansu significantly suppresses thymidylate synthase(TYMS),the enzyme which provides the sole de novo source of thymidylate for DNA synthesis.The inhibition of TYMS could lead to cell-cycle block and DNA damage of HCC cells.Furthermore,we identified that Huachansu markedly increased y-H2AX expression,which indicated the presence of DNA damage.Moreover,we confirmed that transfection of cells with small interfering RNA specific to TYMS could increase the suppressive effects of Huachansu on the HCC cells proliferation.Quantitative RT-PCR analysis showed that Huachansu treatment had no effect on the transcription level of TYMS.Fu rthermore,proteasomal inhibitor MG132 could block TYMS inhibition induced by Huachansu,and concomita nt administration of protein synthesis inhibitor cycloheximide(CHX)with Huachansu could further suppress the protein level of TYMS,indicating that Huachansu promotes proteasome-dependent degradation of TYMS in liver cancer cells.More importantly,the bioactive bufadienolides of Huachansu such as Bufalin,Bufotalin,Cinobufotalin,Desacetylcinobufagin,Arenobufagin,Telocinobufagin,and Resibufogenin could also significantly restrain the protein level of TYMS,revealing the material basis of inhibition of TYMS exposed to Huachansu.5-Fluorouracil(5-FU)is a TYMS inhibitor,we also evaluate the effects of the combined treatment of Huachansu with 5-FU,the results show that interactions between Huachansu and 5-FU are synergistic or antagonistic.Thus,in clinical,attention should be paid to the dosage of Huachansu in combination with 5-FU.Conclusion:Huachansu inhibits the growth and induces DNA damage of human HCC cells through proteasome-dependent degradation of TYMS,bioactive bufadienolides are the material basis of downregulation of TYMS induced by Huachansu.
