Relict subtropical coniferous forests in China face severe fragmentation,resulting in declining populations,and some are under significant threat from invasive alien species.Despite the crucial importance of understan...Relict subtropical coniferous forests in China face severe fragmentation,resulting in declining populations,and some are under significant threat from invasive alien species.Despite the crucial importance of understanding forest dynamics,knowledge gaps persist,particularly regarding the impact of invasive plants on vulnerable natives like Keteleeria evelyniana.In this study,we investigated the impact of invasive plants on the regeneration of forests dominated by K.evelyniana,a subtropical relict species in southwestern China.For this purpose,we characterized forest dynamics of 160 forest plots featuring K.evelyniana as the primary dominant species and determined whether the presence of invasive plants was correlated with regeneration of K.evelyniana.We identified four distinct forest types in which K.evelyniana was dominant.We found that radial growth of K.evelyniana trees is faster in younger ageclasses today than it was for older trees at the same age.The population structure of K.evelyniana in each forest type exhibited a multimodal age-class distribution.However,three forest types lacked established saplings younger than 10 years old,a situation attributed to the dense coverage of the invasive alien Ageratina adenophora.This invasive species resulted in a reduction of understory species diversity.Additionally,our analysis uncovered a significant negative correlation in phylogenetic relatedness(net relatedness index)between native and invasive alien plant species in eastern Yunnan.This suggests closely related invasive species face heightened competition,hindering successful invasion.Taken together,our findings indicate that successful establishment and habitat restoration of K.evelyniana seedling/saplings require effective measures to control invasive plants.展开更多
Pseudotsuga forrestii is a relict evergreen coniferous tree species in Pinaceae endemic to China.P.forrestii tree numbers have greatly decreased due to deforestation,over-utilization and habitat degradation.Here we cl...Pseudotsuga forrestii is a relict evergreen coniferous tree species in Pinaceae endemic to China.P.forrestii tree numbers have greatly decreased due to deforestation,over-utilization and habitat degradation.Here we clarify P.forrestii community types,structure,species diversity,seedling recruitments and growth trends.We identified four P.forrestii community types:(1)Pseudotsuga forrestii-Quercus guyavifolia-Acer davidii evergreen coniferous and broad-leaved mixed forest;(2)Pseudotsuga forrestii-Pinus yunnanensis-Quercus guyavifolia evergreen coniferous and broad-leaved mixed forest;(3)Pseudotsuga forrestii evergreen coniferous forest;(4)Pseudotsuga forrestii-Abies georgei var.smithii evergreen coniferous forest.P.forrestii forests are characterized by both warm temperate and temperate affinities.Simpson diversity,Pielou evenness,Shannon-Wiener diversity indices ranged from 0.75 to 0.76,0.74-0.81,and 1.62-1.93,respectively,with no significant differences among the four forest types.The forest stratification was multilayered.The canopy layer was generally 10-25 m tall,with the emergent layer reached 25-42 m.DBH and age structures of P.forrestii showed multimodal distributions.Its maximum age P.forrestii was 570 years with a DBH of 143 cm.The growth of annual ring width of P.forrestii was slow,and generally decreased with age,whereas the basal area at the breast height increased with age.Established seedlings/saplings were mainly found in unstable micro-habitats.Regeneration of P.forrestii depends on moderate natural disturbances.Finally,we provide recommendations for P.forrestii conservation.展开更多
This study is to report the clinical experience of of Botulinum toxin type A in combination with robot-assisted training on upper limb spasticity and motor function in stroke patient through a case report.The patient ...This study is to report the clinical experience of of Botulinum toxin type A in combination with robot-assisted training on upper limb spasticity and motor function in stroke patient through a case report.The patient underwent combined treatment with Botulinum toxin type A and robot-assisted training of the upper limb for 3 weeks.Evaluation was performed before and after combined treatment,and 3 months follow-up after discharge.The following outcomes were measured:spasticity by the modified Ashworth scale,pain by the numeric rating scale,motor function by the Fugl-Meyer assessment in upper limb,and activity of daily living by the modified barthel index.The combined Botulinum toxin type A and robot-assisted training treatment had an obvious improvement in upper limb spasticity,pain,motor function and activity of daily living.Botulinum toxin type A combined with robot-assisted training therapy is worthy of further application in patients with spastic stroke.展开更多
AIM： To reveal the expression of multidrug-resistance associated proteins： glutathione-S-transferase π（GSTπ）, P-glycoprotein（P-gp） and vault protein lung resistance protein（LRP） in retinoblastoma（RB） witho...AIM： To reveal the expression of multidrug-resistance associated proteins： glutathione-S-transferase π（GSTπ）, P-glycoprotein（P-gp） and vault protein lung resistance protein（LRP） in retinoblastoma（RB） without any conservative treatment before primary enucleation and to correlate this expression with histopathological tumor features. METHODS： A total of 42 specimens of RB undergone primary enucleation were selected for the research. Sections from the formalin-fixed, paraffin-embedded specimens were stained with HE and immunohistochemistry to detect the expression of GSTπ, P-gp and LRP.RESULTS： GSTπ was expressed in 39/42（92.86%） RBs and in 9/9（100%） well-differentiated RBs. P-gp/GSTπ was found in 30（71.42%） of 42 RBs. Totally 9（21.43%） tumors were well differentiated and 33（78.57%） were poorly differentiated. Totally 15（35.71%） eyes had optic nerve（ON） tumor invasion, 36（85.71%） had choroidal tumor invasion, and 14（33.33%） had simultaneous choroidal and ON invasion. There was no statistically significant relationship between P-gp, GSTπ, LRP positivity and the degree of ocular layer tumor invasion and ON tumor invasion（P〉0.05）. CONCLUSION： RB intrinsically expresses GSTπ, P-gp and LRP. GSTπ expression is positive in 100% welldifferentiation ones, so in which way it is correlated with differentiation. But the other two proteins expressions are not related to tumor differentiation and to the degree of tumor invasion. GSTπ may be a new target of chemotherapy in RB.展开更多
·AIM: To discuss the prognostic significant of autophagy related proteins(ARPs) in retinoblastoma(RB)and to find the molecular marker to distinguish retinocytoma(RC) and RB by investigating the different expressi...·AIM: To discuss the prognostic significant of autophagy related proteins(ARPs) in retinoblastoma(RB)and to find the molecular marker to distinguish retinocytoma(RC) and RB by investigating the different expression profiling of microtubule-associated protein light chain 3(LC3B) and other ARPs in RC and RB.·METHODS: Specimens with retinocytoma region(RCR)or mainly composed with Flexner-Winterstein rosettes(FWR) were screen out from 219 paraffin-embedded RB samples and respectively taken as RCR group and FWR group. Others were taken as undifferentiated(UD) group.Immunochemistry(IHC) of LC3 B and electronic microscopy was used to identify autophagy. The IHC scores of LC3 B and other ARPs, such as Beclin, PTEN,p27, p16INK4 a, mTOR and BCL-2 were compared and correlation analysis was applied to find potential proteins which may involve in autophagy regulation. The prognostics significance of LC3 B was evaluated by comparing the high risk features(HRFs) in 3 groups of total 219 samples.·RESULTS: Twenty-one specimens with RCR and 36 specimens mainly composed with FWR were screen out.RCR cell had a high level of LC3 B and lots of autophagic vacuoles. Beclin, PTEN, p27 had positive correlation with LC3, and p16INK4 ahad negative correlation, while the expression of mTOR and BCL-2 in RCR and RB region did not show any difference. Cases with RCR had lower rate of HRFs than undifferentiated cases.·CONCLUSION: ARPs had different expression pattern between RCR and other pathological types of RB, and could be ideal markers to distinguish RC from RB. Our finding indicated cases with RCR had favorable prognosis just like those with FWR.展开更多
Targeting-induced local lesions in genomes(TILLING)is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants.Although TILLING has been developed for many diploid plants...Targeting-induced local lesions in genomes(TILLING)is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants.Although TILLING has been developed for many diploid plants,the technology has been used in very few polyploid species due to their genomic complexity.Here,we established an efficient capillary electrophoresis-based TILLING platform for allotetraploid cultivated tobacco(Nicotiana tabacum L.)using an ethyl methanesulfonate(EMS)-mutagenized population of 1,536 individuals.We optimized the procedures for endonuclease preparation,leaf tissue sampling,DNA extraction,normalization,pooling,PCR amplification,heteroduplex formation,and capillary electrophoresis.In a test screen using seven target genes with eight PCR fragments,we obtained 118 mutants.The mutation density was estimated to be approximately one mutation per 106kb on average.Phenotypic analyses showed that mutations in two heavy metal transporter genes,HMA2S and HMA4T,led to reduced accumulation of cadmium and zinc,which was confirmed independently using CRISPR/Cas9 to generate knockout mutants.Our results demonstrate that this powerful TILLING platform(available at http://www.croptilling.org)can be used in tobacco to facilitate functional genomics applications.展开更多
Diversity arrays technology （DART） is a microarray-based marker system that achieves high throughput by reducing the complexity of the genome. A DArT chip has recently been developed for tobacco. In this study, we g...Diversity arrays technology （DART） is a microarray-based marker system that achieves high throughput by reducing the complexity of the genome. A DArT chip has recently been developed for tobacco. In this study, we genotyped 267 flue-cured cultivars/landraces, including 121 Chinese accessions over five decades from widespread geographic regions in China, 103 from the Americas, and 43 other foreign cultivars, using the newly developed chip. Three hundred and thirty polymorphic DArT makers were selected and used for a phylogenetic analysis, which sug- gested that the 267 accessions could be classified into two subgroups, which could each be further divided into 2-4 sections. Eight elite cultivars, which account for 83% of the area of Chinese tobacco production, were all found in one subgroup. Two high-quality cultivars, HHDJY and Cuibil, were grouped together in one section, while six other high-yield cultivars were grouped into another section. The 330 DArT marker clones were sequenced and close to 95% of them are within non-repetitive regions. Finally, the implications of this study for Chinese flue-cured tobacco breeding and production programs were discussed.展开更多
基金funded by the Major Program for Basic Research Project of Yunnan Province,China,grant number 202101BC070002the Special Foundation for National Science and Technology Basic Resources Investigation of China,grant number 2019FY202300.
文摘Relict subtropical coniferous forests in China face severe fragmentation,resulting in declining populations,and some are under significant threat from invasive alien species.Despite the crucial importance of understanding forest dynamics,knowledge gaps persist,particularly regarding the impact of invasive plants on vulnerable natives like Keteleeria evelyniana.In this study,we investigated the impact of invasive plants on the regeneration of forests dominated by K.evelyniana,a subtropical relict species in southwestern China.For this purpose,we characterized forest dynamics of 160 forest plots featuring K.evelyniana as the primary dominant species and determined whether the presence of invasive plants was correlated with regeneration of K.evelyniana.We identified four distinct forest types in which K.evelyniana was dominant.We found that radial growth of K.evelyniana trees is faster in younger ageclasses today than it was for older trees at the same age.The population structure of K.evelyniana in each forest type exhibited a multimodal age-class distribution.However,three forest types lacked established saplings younger than 10 years old,a situation attributed to the dense coverage of the invasive alien Ageratina adenophora.This invasive species resulted in a reduction of understory species diversity.Additionally,our analysis uncovered a significant negative correlation in phylogenetic relatedness(net relatedness index)between native and invasive alien plant species in eastern Yunnan.This suggests closely related invasive species face heightened competition,hindering successful invasion.Taken together,our findings indicate that successful establishment and habitat restoration of K.evelyniana seedling/saplings require effective measures to control invasive plants.
基金This study received financial support from the Science and Technology Department of Yunnan University,China(2019YNU002)Major Program for Basic Research Project of Yunnan Province,China(202101BC070002)the Special Foundation for National Science and Technology Basic Resources Investigation of China(2019FY202300).
文摘Pseudotsuga forrestii is a relict evergreen coniferous tree species in Pinaceae endemic to China.P.forrestii tree numbers have greatly decreased due to deforestation,over-utilization and habitat degradation.Here we clarify P.forrestii community types,structure,species diversity,seedling recruitments and growth trends.We identified four P.forrestii community types:(1)Pseudotsuga forrestii-Quercus guyavifolia-Acer davidii evergreen coniferous and broad-leaved mixed forest;(2)Pseudotsuga forrestii-Pinus yunnanensis-Quercus guyavifolia evergreen coniferous and broad-leaved mixed forest;(3)Pseudotsuga forrestii evergreen coniferous forest;(4)Pseudotsuga forrestii-Abies georgei var.smithii evergreen coniferous forest.P.forrestii forests are characterized by both warm temperate and temperate affinities.Simpson diversity,Pielou evenness,Shannon-Wiener diversity indices ranged from 0.75 to 0.76,0.74-0.81,and 1.62-1.93,respectively,with no significant differences among the four forest types.The forest stratification was multilayered.The canopy layer was generally 10-25 m tall,with the emergent layer reached 25-42 m.DBH and age structures of P.forrestii showed multimodal distributions.Its maximum age P.forrestii was 570 years with a DBH of 143 cm.The growth of annual ring width of P.forrestii was slow,and generally decreased with age,whereas the basal area at the breast height increased with age.Established seedlings/saplings were mainly found in unstable micro-habitats.Regeneration of P.forrestii depends on moderate natural disturbances.Finally,we provide recommendations for P.forrestii conservation.
基金This study was supported by the grant from the Gansu Provincial Hospital in China(No.18GSSY4-31,No.20GSSY4-52)The authors would like to thank all of the study participants.
文摘This study is to report the clinical experience of of Botulinum toxin type A in combination with robot-assisted training on upper limb spasticity and motor function in stroke patient through a case report.The patient underwent combined treatment with Botulinum toxin type A and robot-assisted training of the upper limb for 3 weeks.Evaluation was performed before and after combined treatment,and 3 months follow-up after discharge.The following outcomes were measured:spasticity by the modified Ashworth scale,pain by the numeric rating scale,motor function by the Fugl-Meyer assessment in upper limb,and activity of daily living by the modified barthel index.The combined Botulinum toxin type A and robot-assisted training treatment had an obvious improvement in upper limb spasticity,pain,motor function and activity of daily living.Botulinum toxin type A combined with robot-assisted training therapy is worthy of further application in patients with spastic stroke.
基金Supported by the National Natural Science Foundation of China(No.30371515)
文摘AIM： To reveal the expression of multidrug-resistance associated proteins： glutathione-S-transferase π（GSTπ）, P-glycoprotein（P-gp） and vault protein lung resistance protein（LRP） in retinoblastoma（RB） without any conservative treatment before primary enucleation and to correlate this expression with histopathological tumor features. METHODS： A total of 42 specimens of RB undergone primary enucleation were selected for the research. Sections from the formalin-fixed, paraffin-embedded specimens were stained with HE and immunohistochemistry to detect the expression of GSTπ, P-gp and LRP.RESULTS： GSTπ was expressed in 39/42（92.86%） RBs and in 9/9（100%） well-differentiated RBs. P-gp/GSTπ was found in 30（71.42%） of 42 RBs. Totally 9（21.43%） tumors were well differentiated and 33（78.57%） were poorly differentiated. Totally 15（35.71%） eyes had optic nerve（ON） tumor invasion, 36（85.71%） had choroidal tumor invasion, and 14（33.33%） had simultaneous choroidal and ON invasion. There was no statistically significant relationship between P-gp, GSTπ, LRP positivity and the degree of ocular layer tumor invasion and ON tumor invasion（P〉0.05）. CONCLUSION： RB intrinsically expresses GSTπ, P-gp and LRP. GSTπ expression is positive in 100% welldifferentiation ones, so in which way it is correlated with differentiation. But the other two proteins expressions are not related to tumor differentiation and to the degree of tumor invasion. GSTπ may be a new target of chemotherapy in RB.
基金Supported by National Natural Science Foundation of China(No.30672276)
文摘·AIM: To discuss the prognostic significant of autophagy related proteins(ARPs) in retinoblastoma(RB)and to find the molecular marker to distinguish retinocytoma(RC) and RB by investigating the different expression profiling of microtubule-associated protein light chain 3(LC3B) and other ARPs in RC and RB.·METHODS: Specimens with retinocytoma region(RCR)or mainly composed with Flexner-Winterstein rosettes(FWR) were screen out from 219 paraffin-embedded RB samples and respectively taken as RCR group and FWR group. Others were taken as undifferentiated(UD) group.Immunochemistry(IHC) of LC3 B and electronic microscopy was used to identify autophagy. The IHC scores of LC3 B and other ARPs, such as Beclin, PTEN,p27, p16INK4 a, mTOR and BCL-2 were compared and correlation analysis was applied to find potential proteins which may involve in autophagy regulation. The prognostics significance of LC3 B was evaluated by comparing the high risk features(HRFs) in 3 groups of total 219 samples.·RESULTS: Twenty-one specimens with RCR and 36 specimens mainly composed with FWR were screen out.RCR cell had a high level of LC3 B and lots of autophagic vacuoles. Beclin, PTEN, p27 had positive correlation with LC3, and p16INK4 ahad negative correlation, while the expression of mTOR and BCL-2 in RCR and RB region did not show any difference. Cases with RCR had lower rate of HRFs than undifferentiated cases.·CONCLUSION: ARPs had different expression pattern between RCR and other pathological types of RB, and could be ideal markers to distinguish RC from RB. Our finding indicated cases with RCR had favorable prognosis just like those with FWR.
基金supported by Yunnan Province Tobacco funding(2014YN05,2014YN06,2016YN25,2017YN03)Bureau of National Tobacco funding(110201401006[JY-06],110201601031[JY-05],110201401005[JY-05]).
文摘Targeting-induced local lesions in genomes(TILLING)is a powerful reverse-genetics tool that enables high-throughput screening of genomic variations in plants.Although TILLING has been developed for many diploid plants,the technology has been used in very few polyploid species due to their genomic complexity.Here,we established an efficient capillary electrophoresis-based TILLING platform for allotetraploid cultivated tobacco(Nicotiana tabacum L.)using an ethyl methanesulfonate(EMS)-mutagenized population of 1,536 individuals.We optimized the procedures for endonuclease preparation,leaf tissue sampling,DNA extraction,normalization,pooling,PCR amplification,heteroduplex formation,and capillary electrophoresis.In a test screen using seven target genes with eight PCR fragments,we obtained 118 mutants.The mutation density was estimated to be approximately one mutation per 106kb on average.Phenotypic analyses showed that mutations in two heavy metal transporter genes,HMA2S and HMA4T,led to reduced accumulation of cadmium and zinc,which was confirmed independently using CRISPR/Cas9 to generate knockout mutants.Our results demonstrate that this powerful TILLING platform(available at http://www.croptilling.org)can be used in tobacco to facilitate functional genomics applications.
基金supported by the China National Tobacco Corporation (Nos. 110200701023 and 110201101010-JY-04)the Yunnan Provincial Tobacco Company (Nos. 08A05 and 2010YN02), China
文摘Diversity arrays technology （DART） is a microarray-based marker system that achieves high throughput by reducing the complexity of the genome. A DArT chip has recently been developed for tobacco. In this study, we genotyped 267 flue-cured cultivars/landraces, including 121 Chinese accessions over five decades from widespread geographic regions in China, 103 from the Americas, and 43 other foreign cultivars, using the newly developed chip. Three hundred and thirty polymorphic DArT makers were selected and used for a phylogenetic analysis, which sug- gested that the 267 accessions could be classified into two subgroups, which could each be further divided into 2-4 sections. Eight elite cultivars, which account for 83% of the area of Chinese tobacco production, were all found in one subgroup. Two high-quality cultivars, HHDJY and Cuibil, were grouped together in one section, while six other high-yield cultivars were grouped into another section. The 330 DArT marker clones were sequenced and close to 95% of them are within non-repetitive regions. Finally, the implications of this study for Chinese flue-cured tobacco breeding and production programs were discussed.
