ABC immunoperoxidase was used to test the effects of rhTGF-β1 and rhGM-CSF on receptor expressions in J6-1 and J6-2 leukemic cell lines. Computer assisted image analysis system was introduced to evaluate positive ind...ABC immunoperoxidase was used to test the effects of rhTGF-β1 and rhGM-CSF on receptor expressions in J6-1 and J6-2 leukemic cell lines. Computer assisted image analysis system was introduced to evaluate positive index of time-and dose-dependent specimens. The expression of c-kit was elevated both in positive rate and positive index by TGF-01 in both time- and dose-dependent manners. Ing/ml rhTGF-β1 simultaneously enhanced the expression of c-fms and PDGF-R which is not detected in 50 ng / ml GM-CSF treatment. Endoglin was down-regulated after TGF-β treatment and up-regulated in J6-2 cells after GM-CSF treatment, c-kit Expression was elevated by TGF-β in J6-1 cells while decreased by both in J6-2 cells.展开更多
It’s very necessary to study the translation of figurative expressions from English into Chinese since two languages differ a lot in lexical,syntactic and cultural aspects.In terms of translation approaches there are...It’s very necessary to study the translation of figurative expressions from English into Chinese since two languages differ a lot in lexical,syntactic and cultural aspects.In terms of translation approaches there are three ones largely used depending on different situations: literal translation from English into Chinese,translation from figurative expressions into figurative ones and translation from figurative expressions into nonfigurative ones.展开更多
Background： The SWEET （Sugars will eventually be exported transporters） gene family plays multiple roles in plant physiological activities and development process. It participates in reproductive development and in...Background： The SWEET （Sugars will eventually be exported transporters） gene family plays multiple roles in plant physiological activities and development process. It participates in reproductive development and in the process of sugar transport and absorption, plant senescence and stress responses and plant-pathogen interaction. However, thecomprehensive analysis of SWEET genes has not been reported in cotton. Results： In this study, we identified 22, 31, 55 and 60 SWEETgenes from the sequenced genomes of Gossypium orboreum, G. rairnondii, G. hirsutum and G. borbadense, respectively. Phylogenetic tree analysis showed that the SWEET genes could be divided into four groups, which were further classified into 14 sub-clades. Further analysis of chromosomal location, synteny analysis and gene duplication suggested that the orthologs showed a good collinearity and segmental duplication events played a crucial role in the expansion of the family in cotton. Specific MtN3_slv domains were highly conserved between Arabidopsis and cotton by exon-intron organization and motif analysis. In addition, the expression pattern in different tissues indicated that the duplicated genes in cotton might have acquired new functions as a result of sub-functionalization or neo-functionalization. The expression pattern of SWEET genes showed that the different genes were induced by diverse stresses. The identification and functional analysis of SWEET genes in cotton may provide more candidate genes for genetic modification. Conclusion： SWEET genes were classified into four clades in cotton. The expression patterns suggested that the duplicated genes might have experienced a functional divergence. This work provides insights into the evolution of SWEETgenes and more candidates for specific genetic modification, which will be useful in future research.展开更多
The fruit of Amomum villosum is highly valued for its medicinal and edible properties.However,the fruit abscission rate is exceptionally high,resulting in low unit yield.To better understand the fruit growth and devel...The fruit of Amomum villosum is highly valued for its medicinal and edible properties.However,the fruit abscission rate is exceptionally high,resulting in low unit yield.To better understand the fruit growth and development,this study investigated the characteristic changes of fruits and the expression patterns of GA-related genes in both fruits and fruit stalks.Results revealed that it takes approximately 90 days for the ovary of A.villosum to reach maturity.Fruit growth pattern exhibited a slow-fast-slow trend,with the peak of fruit weight accumulation occurring between the 14 and 25 days after artificial pollination(DAP).Before the 27 DAP,significant changes were observed in the ovary,fruit color,pericarp thickness,and fruit thorn length.Additionally,ten candidate GA-related genes,including 1 GA_(13)ox,1 GA_(20)ox,1 GA_(3)ox,2 GA_(2)oxs,4 GID1cs,and 1 DELLA,were analyzed using bioinformatics tools.The GA-related genes in fruit and fruit stalk exhibited significant correlations with fruit growth/development and fruit dropping.Overall,the fruit growth and development law,and the expression patterns of GA-related genes provided a theoretical foundation for further functional study and the screening of candidate genes for potential applications in A.villosum.展开更多
Dianthus spiculifolius Schur,as an emerging ornamental plant,has extensive applications and economic values.In this study,the DsCBL4 gene was successfully cloned,and its tissue-specific expression,expression patterns ...Dianthus spiculifolius Schur,as an emerging ornamental plant,has extensive applications and economic values.In this study,the DsCBL4 gene was successfully cloned,and its tissue-specific expression,expression patterns under various abiotic stresses,subcellular localization,and bioinformatics analysis of the encoded amino acid sequence were conducted.The results showed that the coding region of the DsCBL4 gene was 675 bp long,encoding 224 amino acids.It had high homology with the amino acids encoded by Amaranthus tricolor,Chenopodium quinoa and Spinacia oleracea.The predicted relative molecular mass of DsCBL4 was 25.61 ku,with an isoelectric point of 4.58,and it had phosphorylation sites,belonging to an unstable hydrophilic protein.Its secondary structure includedα-helices,irregular coils and extended chains.The tertiary structure prediction revealed that DsCBL4 had four EFhand calcium-binding domains necessary for Ca2+binding in plant calmodulin-like proteins and the FPSF motif for calcineurin B-like protein(CBL)-interacting protein kinase(CIPK)activation.The expression level of the DsCBL4 gene showed tissue specificity,with the highest expression in roots.It was induced by drought,low temperature,combined drought and low temperature,salt stress,nitrogen stress,phosphorus stress,calcium ion stress,high temperature stress,and abscisic acid(ABA)stress.Both transient infection in Nicotiana tabacum L.and stable expression in transgenic Arabidopsis thaliana showed that the DsCBL4 protein was localized to the cell membrane.These results suggested that DsCBL4 might be involved in the abiotic stress response of Dianthus spiculifolius through the calcium signaling pathway,providing a theoretical basis for understanding its molecular mechanism.This study provided an important reference for further exploring the role of the DsCBLs gene family in plant stress resistance.展开更多
Objective INF2 is a member of the formins family.Abnormal expression and regulation of INF2 have been associated with the progression of various tumors,but the expression and role of INF2 in hepatocellular carcinoma(H...Objective INF2 is a member of the formins family.Abnormal expression and regulation of INF2 have been associated with the progression of various tumors,but the expression and role of INF2 in hepatocellular carcinoma(HCC)remain unclear.HCC is a highly lethal malignant tumor.Given the limitations of traditional treatments,this study explored the expression level,clinical value and potential mechanism of INF2 in HCC in order to seek new therapeutic targets.Methods In this study,we used public databases to analyze the expression of INF2 in pan-cancer and HCC,as well as the impact of INF2 expression levels on HCC prognosis.Quantitative real time polymerase chain reaction(RT-qPCR),Western blot,and immunohistochemistry were used to detect the expression level of INF2 in liver cancer cells and human HCC tissues.The correlation between INF2 expression and clinical pathological features was analyzed using public databases and clinical data of human HCC samples.Subsequently,the effects of INF2 expression on the biological function and Drp1 phosphorylation of liver cancer cells were elucidated through in vitro and in vivo experiments.Finally,the predictive value and potential mechanism of INF2 in HCC were further analyzed through database and immunohistochemical experiments.Results INF2 is aberrantly high expression in HCC samples and the high expression of INF2 is correlated with overall survival,liver cirrhosis and pathological differentiation of HCC patients.The expression level of INF2 has certain diagnostic value in predicting the prognosis and pathological differentiation of HCC.In vivo and in vitro HCC models,upregulated expression of INF2 triggers the proliferation and migration of the HCC cell,while knockdown of INF2 could counteract this effect.INF2 in liver cancer cells may affect mitochondrial division by inducing Drp1 phosphorylation and mediate immune escape by up-regulating PD-L1 expression,thus promoting tumor progression.Conclusion INF2 is highly expressed in HCC and is associated with poor prognosis.High expression of INF2 may promote HCC progression by inducing Drp1 phosphorylation and up-regulation of PD-L1 expression,and targeting INF2 may be beneficial for HCC patients with high expression of INF2.展开更多
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
Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and ...Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and hepatocyte ballooning,and may further progress to liver fibrosis and cirrhosis.Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),a member of the scavenger receptor family,recognizes and binds oxidized low-density lipoprotein.This study aims to investigate the role of LOX-1 in MASH progression.Methods:LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus(GEO)datasets.Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin(α-SMA)levels and co-localization in fibrotic liver tissues and LX-2 cells.LOX-1 knockout(Lox-1^(−/−))mice were generated using CRISPR/caspase-9(Cas9)and genotyped by PCR and Sanger sequencing.Wild-type(WT)and Lox-1^(−/−)mice were randomized into control and Western diet model groups.Serum and liver samples were collected for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)measurement by biochemical kits,liver structure evaluation by hematoxylin and eosin(HE)staining,collagen deposition by Masson staining,lipid accumulation by Oil Red O staining,and fibrotic marker gene expression by real-time quantitative PCR(RT-qPCR).Network pharmacology and search tool for the retrieval of interacting genes/proteins(STRING)-based protein-protein interaction(PPI)with Gene Ontology(GO)enrichment were used to predict downstream targets and pathways.Results:The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet(HFD)and bile duct ligation(BDL)mouse models(both P<0.001).LOX-1 and α-SMA levels were elevated in fibrotic liver tissues.Lox-1^(−/−)mice were successfully established.Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet(both P<0.001),and these levels decreased after LOX-1 knockout(both P<0.05).HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration,steatosis,inflammatory cell infiltration,and periportal fibroplasia,which were significantly ameliorated by LOX-1 knockout.Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with controldiet mice,and LOX-1 knockout inhibited collagen fiber deposition(all P<0.05).RT‑qPCR results showed that hepatic mRNA levels of Acta2,Col1a1,and Timp1 were significantly increased in Western diet-fed mice,and LOX-1 knockout reduced the expression of these fibrogenic marker genes.Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged,displayed macrovesicular steatosis,and exhibited diffusely distributed red lipid droplets,whereas LOX-1 knockout alleviated hepatic lipid accumulation(both P<0.001).RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2,Col1a1,and Timp1 mRNA levels in LX‑2 cells(all P<0.05).Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells,and LOX-1 silencing suppressed α‑SMA expression.Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks.Conclusion:LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.展开更多
Background Cotton is an industrial crop renowned for its multifaceted applications in the textiles,pharmaceuticals,and biofuel industries.Plant regeneration through somatic embryogenesis(SE)plays a crucial role in the...Background Cotton is an industrial crop renowned for its multifaceted applications in the textiles,pharmaceuticals,and biofuel industries.Plant regeneration through somatic embryogenesis(SE)plays a crucial role in the genetic improvement of cotton.There is a strong correlation between SE and zygotic embryogenesis(ZE)in plants.Furthermore,the strategy of ectopic expression of cotton genes into the model plant Arabidopsis has been a widely accepted approach for functional study.Result Based on previous spatial transcriptomics of cotton somatic embryos,two genes,Gh HAT5 and Gh CRK29,were identified.They are highly expressed in cotyledon and epidermal cells of cotton cotyledonary embryos,respectively.In this study,Gh HAT5 and Gh CRK29 were ectopically expressed in Arabidopsis to investigate their functions.The result showed that in Arabidopsis zygotic embryos,the overexpression of Gh HAT5 promoted the development of apical embryonic upper-tier cells and embryonic cotyledon,while the overexpression of Gh CRK29 promoted the development of apical embryonic lower-tier cells and embryonic radicle.Given the similarities between somatic and zygotic embryogenesis,these findings suggest that Gh HAT5 and Gh CRK29 are involved in cotton SE.We also speculate that these genes may promote the expression of the Arabidopsis endogenous gene At SCR,which is crucial for embryonic development.Conclusion These results revealed that Gh HAT5 and Gh CRK29 regulate embryonic development and are essential in advancing our understanding of cotton SE and facilitating targeted genetic manipulation strategies to improve industrial crop traits and agricultural sustainability.展开更多
Background Cotton is an important crop providing the most natural fibers all over the world. The cotton genomics community has utilized whole genome sequencing data to construct an elite gene pool in which functional ...Background Cotton is an important crop providing the most natural fibers all over the world. The cotton genomics community has utilized whole genome sequencing data to construct an elite gene pool in which functional genes are related to agronomic traits. However, the functional validation of these genes is hindered by time-consuming and inefficient genetic transformation methods. Thus, establishing a transient transformation system of high efficiency is necessary for cotton genomics.Results To improve the efficiency of transient transformation, we used the protoplasts isolated from the etiolated cotyledon as recipient. The enzymatic digestion buffer comprised 1.5%(w/v) cellulase, 0.75%(w/v) macerozyme, and 1% hemicellulase, osmotically buffered with 0.4 mol·L^(-1) mannitol. After 5 h of dark incubation at 25℃, uniform cotton protoplasts were successfully isolated with a yield of 4.6 × 10^(6) protoplasts per gram(fresh weight) and 95% viability. We incubated 100 μL protoplasts(2.5 × 10^(5)·m L^(-1)) with 15 μg plasmid in the solution of 0.4 mol·L^(-1) mannitol and 40% PEG 4000 for 15 min, ultimately achieving an optimal transient transfection efficiency of 71.47%.Conclusions This transient system demonstrated effective utility in cellular biology research through successful applications in subcellular localization analyses, bimolecular fluorescence complementation(Bi FC) verification, and prime editing vector validation. Through systematic optimization, we established an efficient and expedited protoplast-based transient transformation system and successfully applied this platform to cotton functional genomics studies.展开更多
As a medium between two different languages, between peoples of two countries, translation is not only a matter of language, but also of cross-cultural transference. According to Professor He Ziran, socio-pragmatic tr...As a medium between two different languages, between peoples of two countries, translation is not only a matter of language, but also of cross-cultural transference. According to Professor He Ziran, socio-pragmatic translation is the kind of translation which examines the conditions on language use that stem from the social and cultural situations to serve cross-cultural communication. This paper focuses on the ways used to achieve socio-pragmatic equivalence in translation practice.展开更多
A pair of specific primers were designed and synthesized according to the published sequences of ORF1 gene of PCV 2. The complete DNA fragment of ORF1 gene was obtained by PCR from viral DNA of PCV 2 QD strain.Its nuc...A pair of specific primers were designed and synthesized according to the published sequences of ORF1 gene of PCV 2. The complete DNA fragment of ORF1 gene was obtained by PCR from viral DNA of PCV 2 QD strain.Its nuclectide sequence was determined by the dideoxy mediated chain termination method.The results showed that the complete open reading frame (ORF) of ORF1 gene encoding 314 amino acids was 945 bp in length.A comparison of the nucleotide and amino acid sequences of ORF1 gene with that of other PCV strains showed that the identity of nucleotide with PCV 1 and PCV 2 were 83% and 96 4%～99 2% respectively,and identity of the deduced amino acid with PCV 1 and PCV 2 were 84% and more than 98% respecitively.The DNA fragment of ORF1 gene was subcloned into prokaryotic expression vector pET 28a and pGEX KG;while the specific non fusion and fusion proteins with GST of molecular weight 38 kD and 63 kD were expressed in E.coli BL 21 (DE3).Western blotting assay indicated that the polyclonal antibody against PCV 2 could recognize these two proteins.展开更多
基金Supported by National Natural Sciences Foundation. The abstract of this work was published in Exp Hematol (1994:22:743)
文摘ABC immunoperoxidase was used to test the effects of rhTGF-β1 and rhGM-CSF on receptor expressions in J6-1 and J6-2 leukemic cell lines. Computer assisted image analysis system was introduced to evaluate positive index of time-and dose-dependent specimens. The expression of c-kit was elevated both in positive rate and positive index by TGF-01 in both time- and dose-dependent manners. Ing/ml rhTGF-β1 simultaneously enhanced the expression of c-fms and PDGF-R which is not detected in 50 ng / ml GM-CSF treatment. Endoglin was down-regulated after TGF-β treatment and up-regulated in J6-2 cells after GM-CSF treatment, c-kit Expression was elevated by TGF-β in J6-1 cells while decreased by both in J6-2 cells.
文摘It’s very necessary to study the translation of figurative expressions from English into Chinese since two languages differ a lot in lexical,syntactic and cultural aspects.In terms of translation approaches there are three ones largely used depending on different situations: literal translation from English into Chinese,translation from figurative expressions into figurative ones and translation from figurative expressions into nonfigurative ones.
基金supported by the The National Key ResearchDevelopment Program of China(2016YFD0101400,2017YFD0101600)
文摘Background： The SWEET （Sugars will eventually be exported transporters） gene family plays multiple roles in plant physiological activities and development process. It participates in reproductive development and in the process of sugar transport and absorption, plant senescence and stress responses and plant-pathogen interaction. However, thecomprehensive analysis of SWEET genes has not been reported in cotton. Results： In this study, we identified 22, 31, 55 and 60 SWEETgenes from the sequenced genomes of Gossypium orboreum, G. rairnondii, G. hirsutum and G. borbadense, respectively. Phylogenetic tree analysis showed that the SWEET genes could be divided into four groups, which were further classified into 14 sub-clades. Further analysis of chromosomal location, synteny analysis and gene duplication suggested that the orthologs showed a good collinearity and segmental duplication events played a crucial role in the expansion of the family in cotton. Specific MtN3_slv domains were highly conserved between Arabidopsis and cotton by exon-intron organization and motif analysis. In addition, the expression pattern in different tissues indicated that the duplicated genes in cotton might have acquired new functions as a result of sub-functionalization or neo-functionalization. The expression pattern of SWEET genes showed that the different genes were induced by diverse stresses. The identification and functional analysis of SWEET genes in cotton may provide more candidate genes for genetic modification. Conclusion： SWEET genes were classified into four clades in cotton. The expression patterns suggested that the duplicated genes might have experienced a functional divergence. This work provides insights into the evolution of SWEETgenes and more candidates for specific genetic modification, which will be useful in future research.
基金Supported by Seed Industry Vitalization Project of Rural Revitalization Strategy of Guangdong Province of China(2024-440000-90060000-8796)。
文摘The fruit of Amomum villosum is highly valued for its medicinal and edible properties.However,the fruit abscission rate is exceptionally high,resulting in low unit yield.To better understand the fruit growth and development,this study investigated the characteristic changes of fruits and the expression patterns of GA-related genes in both fruits and fruit stalks.Results revealed that it takes approximately 90 days for the ovary of A.villosum to reach maturity.Fruit growth pattern exhibited a slow-fast-slow trend,with the peak of fruit weight accumulation occurring between the 14 and 25 days after artificial pollination(DAP).Before the 27 DAP,significant changes were observed in the ovary,fruit color,pericarp thickness,and fruit thorn length.Additionally,ten candidate GA-related genes,including 1 GA_(13)ox,1 GA_(20)ox,1 GA_(3)ox,2 GA_(2)oxs,4 GID1cs,and 1 DELLA,were analyzed using bioinformatics tools.The GA-related genes in fruit and fruit stalk exhibited significant correlations with fruit growth/development and fruit dropping.Overall,the fruit growth and development law,and the expression patterns of GA-related genes provided a theoretical foundation for further functional study and the screening of candidate genes for potential applications in A.villosum.
基金Supported by the National Key R&D Program(2022YFF1300500)the Spring Goose Support Program(CYQN24018)the National Natural Science Foundation of China(32572123)。
文摘Dianthus spiculifolius Schur,as an emerging ornamental plant,has extensive applications and economic values.In this study,the DsCBL4 gene was successfully cloned,and its tissue-specific expression,expression patterns under various abiotic stresses,subcellular localization,and bioinformatics analysis of the encoded amino acid sequence were conducted.The results showed that the coding region of the DsCBL4 gene was 675 bp long,encoding 224 amino acids.It had high homology with the amino acids encoded by Amaranthus tricolor,Chenopodium quinoa and Spinacia oleracea.The predicted relative molecular mass of DsCBL4 was 25.61 ku,with an isoelectric point of 4.58,and it had phosphorylation sites,belonging to an unstable hydrophilic protein.Its secondary structure includedα-helices,irregular coils and extended chains.The tertiary structure prediction revealed that DsCBL4 had four EFhand calcium-binding domains necessary for Ca2+binding in plant calmodulin-like proteins and the FPSF motif for calcineurin B-like protein(CBL)-interacting protein kinase(CIPK)activation.The expression level of the DsCBL4 gene showed tissue specificity,with the highest expression in roots.It was induced by drought,low temperature,combined drought and low temperature,salt stress,nitrogen stress,phosphorus stress,calcium ion stress,high temperature stress,and abscisic acid(ABA)stress.Both transient infection in Nicotiana tabacum L.and stable expression in transgenic Arabidopsis thaliana showed that the DsCBL4 protein was localized to the cell membrane.These results suggested that DsCBL4 might be involved in the abiotic stress response of Dianthus spiculifolius through the calcium signaling pathway,providing a theoretical basis for understanding its molecular mechanism.This study provided an important reference for further exploring the role of the DsCBLs gene family in plant stress resistance.
文摘Objective INF2 is a member of the formins family.Abnormal expression and regulation of INF2 have been associated with the progression of various tumors,but the expression and role of INF2 in hepatocellular carcinoma(HCC)remain unclear.HCC is a highly lethal malignant tumor.Given the limitations of traditional treatments,this study explored the expression level,clinical value and potential mechanism of INF2 in HCC in order to seek new therapeutic targets.Methods In this study,we used public databases to analyze the expression of INF2 in pan-cancer and HCC,as well as the impact of INF2 expression levels on HCC prognosis.Quantitative real time polymerase chain reaction(RT-qPCR),Western blot,and immunohistochemistry were used to detect the expression level of INF2 in liver cancer cells and human HCC tissues.The correlation between INF2 expression and clinical pathological features was analyzed using public databases and clinical data of human HCC samples.Subsequently,the effects of INF2 expression on the biological function and Drp1 phosphorylation of liver cancer cells were elucidated through in vitro and in vivo experiments.Finally,the predictive value and potential mechanism of INF2 in HCC were further analyzed through database and immunohistochemical experiments.Results INF2 is aberrantly high expression in HCC samples and the high expression of INF2 is correlated with overall survival,liver cirrhosis and pathological differentiation of HCC patients.The expression level of INF2 has certain diagnostic value in predicting the prognosis and pathological differentiation of HCC.In vivo and in vitro HCC models,upregulated expression of INF2 triggers the proliferation and migration of the HCC cell,while knockdown of INF2 could counteract this effect.INF2 in liver cancer cells may affect mitochondrial division by inducing Drp1 phosphorylation and mediate immune escape by up-regulating PD-L1 expression,thus promoting tumor progression.Conclusion INF2 is highly expressed in HCC and is associated with poor prognosis.High expression of INF2 may promote HCC progression by inducing Drp1 phosphorylation and up-regulation of PD-L1 expression,and targeting INF2 may be beneficial for HCC patients with high expression of INF2.
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.
基金supported by the Natural Science Foundation of Hunan Province,China(211142095031)。
文摘Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and hepatocyte ballooning,and may further progress to liver fibrosis and cirrhosis.Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),a member of the scavenger receptor family,recognizes and binds oxidized low-density lipoprotein.This study aims to investigate the role of LOX-1 in MASH progression.Methods:LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus(GEO)datasets.Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin(α-SMA)levels and co-localization in fibrotic liver tissues and LX-2 cells.LOX-1 knockout(Lox-1^(−/−))mice were generated using CRISPR/caspase-9(Cas9)and genotyped by PCR and Sanger sequencing.Wild-type(WT)and Lox-1^(−/−)mice were randomized into control and Western diet model groups.Serum and liver samples were collected for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)measurement by biochemical kits,liver structure evaluation by hematoxylin and eosin(HE)staining,collagen deposition by Masson staining,lipid accumulation by Oil Red O staining,and fibrotic marker gene expression by real-time quantitative PCR(RT-qPCR).Network pharmacology and search tool for the retrieval of interacting genes/proteins(STRING)-based protein-protein interaction(PPI)with Gene Ontology(GO)enrichment were used to predict downstream targets and pathways.Results:The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet(HFD)and bile duct ligation(BDL)mouse models(both P<0.001).LOX-1 and α-SMA levels were elevated in fibrotic liver tissues.Lox-1^(−/−)mice were successfully established.Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet(both P<0.001),and these levels decreased after LOX-1 knockout(both P<0.05).HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration,steatosis,inflammatory cell infiltration,and periportal fibroplasia,which were significantly ameliorated by LOX-1 knockout.Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with controldiet mice,and LOX-1 knockout inhibited collagen fiber deposition(all P<0.05).RT‑qPCR results showed that hepatic mRNA levels of Acta2,Col1a1,and Timp1 were significantly increased in Western diet-fed mice,and LOX-1 knockout reduced the expression of these fibrogenic marker genes.Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged,displayed macrovesicular steatosis,and exhibited diffusely distributed red lipid droplets,whereas LOX-1 knockout alleviated hepatic lipid accumulation(both P<0.001).RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2,Col1a1,and Timp1 mRNA levels in LX‑2 cells(all P<0.05).Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells,and LOX-1 silencing suppressed α‑SMA expression.Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks.Conclusion:LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.
基金supported by the National Key Research and Development Program of China(No.2022YFD1200300)。
文摘Background Cotton is an industrial crop renowned for its multifaceted applications in the textiles,pharmaceuticals,and biofuel industries.Plant regeneration through somatic embryogenesis(SE)plays a crucial role in the genetic improvement of cotton.There is a strong correlation between SE and zygotic embryogenesis(ZE)in plants.Furthermore,the strategy of ectopic expression of cotton genes into the model plant Arabidopsis has been a widely accepted approach for functional study.Result Based on previous spatial transcriptomics of cotton somatic embryos,two genes,Gh HAT5 and Gh CRK29,were identified.They are highly expressed in cotyledon and epidermal cells of cotton cotyledonary embryos,respectively.In this study,Gh HAT5 and Gh CRK29 were ectopically expressed in Arabidopsis to investigate their functions.The result showed that in Arabidopsis zygotic embryos,the overexpression of Gh HAT5 promoted the development of apical embryonic upper-tier cells and embryonic cotyledon,while the overexpression of Gh CRK29 promoted the development of apical embryonic lower-tier cells and embryonic radicle.Given the similarities between somatic and zygotic embryogenesis,these findings suggest that Gh HAT5 and Gh CRK29 are involved in cotton SE.We also speculate that these genes may promote the expression of the Arabidopsis endogenous gene At SCR,which is crucial for embryonic development.Conclusion These results revealed that Gh HAT5 and Gh CRK29 regulate embryonic development and are essential in advancing our understanding of cotton SE and facilitating targeted genetic manipulation strategies to improve industrial crop traits and agricultural sustainability.
基金supported by Biological Breeding of Early Maturing and Disease Resistant Cotton Varieties (NO.2023ZD04041)the Project of China Agriculture Research System (Grant No. CARS-15-06)+2 种基金Natural Science Foundation of Henan Province (Grant No. 232300421041 and 222300420382)National Natural Science Foundation of China (Grant No. U21 A20213)the Central Public-interest Scientific Institution Basal Research Fund (Grant No. 1610162023017 and 1610162023028)。
文摘Background Cotton is an important crop providing the most natural fibers all over the world. The cotton genomics community has utilized whole genome sequencing data to construct an elite gene pool in which functional genes are related to agronomic traits. However, the functional validation of these genes is hindered by time-consuming and inefficient genetic transformation methods. Thus, establishing a transient transformation system of high efficiency is necessary for cotton genomics.Results To improve the efficiency of transient transformation, we used the protoplasts isolated from the etiolated cotyledon as recipient. The enzymatic digestion buffer comprised 1.5%(w/v) cellulase, 0.75%(w/v) macerozyme, and 1% hemicellulase, osmotically buffered with 0.4 mol·L^(-1) mannitol. After 5 h of dark incubation at 25℃, uniform cotton protoplasts were successfully isolated with a yield of 4.6 × 10^(6) protoplasts per gram(fresh weight) and 95% viability. We incubated 100 μL protoplasts(2.5 × 10^(5)·m L^(-1)) with 15 μg plasmid in the solution of 0.4 mol·L^(-1) mannitol and 40% PEG 4000 for 15 min, ultimately achieving an optimal transient transfection efficiency of 71.47%.Conclusions This transient system demonstrated effective utility in cellular biology research through successful applications in subcellular localization analyses, bimolecular fluorescence complementation(Bi FC) verification, and prime editing vector validation. Through systematic optimization, we established an efficient and expedited protoplast-based transient transformation system and successfully applied this platform to cotton functional genomics studies.
文摘As a medium between two different languages, between peoples of two countries, translation is not only a matter of language, but also of cross-cultural transference. According to Professor He Ziran, socio-pragmatic translation is the kind of translation which examines the conditions on language use that stem from the social and cultural situations to serve cross-cultural communication. This paper focuses on the ways used to achieve socio-pragmatic equivalence in translation practice.
文摘A pair of specific primers were designed and synthesized according to the published sequences of ORF1 gene of PCV 2. The complete DNA fragment of ORF1 gene was obtained by PCR from viral DNA of PCV 2 QD strain.Its nuclectide sequence was determined by the dideoxy mediated chain termination method.The results showed that the complete open reading frame (ORF) of ORF1 gene encoding 314 amino acids was 945 bp in length.A comparison of the nucleotide and amino acid sequences of ORF1 gene with that of other PCV strains showed that the identity of nucleotide with PCV 1 and PCV 2 were 83% and 96 4%～99 2% respectively,and identity of the deduced amino acid with PCV 1 and PCV 2 were 84% and more than 98% respecitively.The DNA fragment of ORF1 gene was subcloned into prokaryotic expression vector pET 28a and pGEX KG;while the specific non fusion and fusion proteins with GST of molecular weight 38 kD and 63 kD were expressed in E.coli BL 21 (DE3).Western blotting assay indicated that the polyclonal antibody against PCV 2 could recognize these two proteins.
