Saline-alkaline stress can dramatically inhibit plant growth and limit crop production. Wild soybean （Glycine soja） is a crop that adapts well to such environmental stresses. In this study, RNA-sequencing technology...Saline-alkaline stress can dramatically inhibit plant growth and limit crop production. Wild soybean （Glycine soja） is a crop that adapts well to such environmental stresses. In this study, RNA-sequencing technology was used to analyze the transcriptome profles of G. soja roots subjected to 50 mmol·L^-1 NaHCO3 and 150 mmol·L^-1 NaCl treatments. Totally, 2 125 differentially-expressed genes （DEGs） after NaCl treatment and 1 839 DEGs after NaHCO3 treatment were identifed. The top 14 DEGs revealed by RNA-seq were analyzed using qRT-PCR （quantitative real-time polymerase chain reaction）. Gene ontology （GO） annotation showed that most of DEGs under salt and alkali stresses were enriched in ＂metabolic process＂, ＂catalytic activity＂ and ＂binding＂ terms. To search for transcription factors （TFs） among DEGs, the data were screened against TF database PlantTFDB, and it was found that fve TF families, Apetala2/ethylene-responsive element binding proteins （AP2-EREBP）, V-myb avian myeloblastosis viral oncogene homolog （MYB）, WRKYGQK and Zinc fnger motif （WRKY）, NAM, ATAF1/2, CUC1/2 （NAC） and Cys2/His2 （C2H2） were involved in salt stress response. Other fve TF families, NAC, WRKY, MYB, AP2-EREBP and bZIP were involved in response to alkali stress. These two stress treatments shared NAC, WRKY, AP2-EREBP and MYB, and the only two different TFs were bZIP and C2H2. Forty-eight MYB TFs were differentially expressed under salt and alkali stresses, and most of them were up-regulated. This study provided useful information for further investigation of DEGs and TFs in response to saline and alkaline stresses and helped in understanding the molecular basis of the response of G. soja to saline and alkaline stresses.展开更多
The outer membrane protein, ompA, ofAeromonas veronii has a role in the virulence of the organism and is a potential candidate for vaccine development. In this study, ompA I ofAeromonas veronii strain WA106 was cloned...The outer membrane protein, ompA, ofAeromonas veronii has a role in the virulence of the organism and is a potential candidate for vaccine development. In this study, ompA I ofAeromonas veronii strain WA106 was cloned and sequenced, then, it was expressed in Escherichia coli BL21. The nucleotide sequence of ompA I gene was 1 023 base pairs （GenBank Accession NO.KC748024）, which showed 100% homology with that of A. veronii （NO.AB290200.1）. This predicted protein was composed of 340 amino acid residues. Its molecular weight was 35.78 ku and isoelectric point was 5.18. The protein was a hydrophilic protein containing alpha helix and random coil with percentage of 35.0% and 49.7%, respectively. The tertiary structure, quaternary structure prediction showed that ompA I protein contained two peptide chains. SDS-PAGE showed that the actual value of the fusion protein was consistent with the expected result. It will facilitate further study of the role of ompA I protein.展开更多
Myoblast differentiation is an essential process during skeletal muscle development.C2C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro.Dehydrogenase/reductase(SDR family...Myoblast differentiation is an essential process during skeletal muscle development.C2C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro.Dehydrogenase/reductase(SDR family)member 3(DHRS3)is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol.Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation.However,the effect of DHRS3 on mouse muscle cell differentiation was unclear.The objective of current study was to determine if DHRS3 affected muscle cell differentiation,and if DHRS3 was involved in muscle regeneration.Protein expression was determined by western blot and immunofluorescence analysis.The activation and inhibition of DHRS3 increased and decreased C2C12 myoblast differentiation respectively,which indicated that DHRS3 could affect C2C12 myoblast differentiation.DHRS3 expression was significantly changed during muscle regeneration,with the regeneration of muscle injury,the expression of DHRS3 tended to increase first and then decrease.It suggested that DHRS3 might be involved in muscle regeneration.In summary,this study confirmed the involvement of DHRS3 in C2C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development.展开更多
In order to obtain pure enzyme with high activity,two amylase producing strains were isolated from soil samples,and named as the strains LZ-10 and LZ-11.According to morphologic observation,physiology and biochemistry...In order to obtain pure enzyme with high activity,two amylase producing strains were isolated from soil samples,and named as the strains LZ-10 and LZ-11.According to morphologic observation,physiology and biochemistry experiments,16S rRNA and gyrB gene analysis,the strains LZ-10 and LZ-11 were identified as Bacillus subtilis.Adopted the method of ammonium sulfate,DEAE-52 anion purify enzyme,finally used polyacrylamide gel electrophoresis(SDS-PAGE)to detect molecular weight.The strain LZ-10 had an amylase activity of 123.3 U·mL^(-1),a purification factor of 6.8,a recovery rate of 69.5% and an optimal temperature of 50℃.The amylase activity of the strain LZ-11 was 59.91 U·mL^(-1),the purification factor was 4.5,the recovery rate was 60.5%,and the optimum temperature was 55℃.The commodity enzyme derived from Bacillus subtilis was 37.5 U·mL^(-1).The relative molecular weight of amylase activity from the two strains was 55 ku.Both thermal stability and pH stability were higher than those of commercialized amylase.展开更多
According to our previous study, saprophytic fungi Botrytis cinerea contained 579 predicted secretary proteins. Among them, we found that 122 of these proteins contained the highly conserved pathogenic-related host-ta...According to our previous study, saprophytic fungi Botrytis cinerea contained 579 predicted secretary proteins. Among them, we found that 122 of these proteins contained the highly conserved pathogenic-related host-targeting-motif RxLx within 100 residues adjacent to the signal peptide cleavage site. According to PEDNAT and COG of the GenBank database, the functions of this motif containing proteins included metabolism modification and cell secretion. We blasted them in GenBank and found 47.54% had highly conserved homologues in other species, among them 74.1% had putative functional domains. This suggests these proteins are presumably ancient and vertically transmitted within the species. Many of these domains belonged to proteins which played roles in the pathogenic process of other kinds of pathogens and some had already been proved to be pathogenic secretary proteins of Botrytis cinerea. So we postulated that proteins contained host-targeting-motif RxLx were candidates participating in the pathogenesis of Botrytis cinerea.展开更多
It is well established that living cells and tissues respond to mechanical forces such as flow-related shear stresses in blood or interstitial space and complex tractional stresses at cell-matrix contacts and cell-cel...It is well established that living cells and tissues respond to mechanical forces such as flow-related shear stresses in blood or interstitial space and complex tractional stresses at cell-matrix contacts and cell-cell contacts.However,how different modes of forces impact mechanical and biological responses is elusive.Here we describe a strategy of using the three-dimensional magnetic twisting cytometry(3D MTC)technology to apply forces in any desired directions to the same living cell.We reveal that for a fixed stress amplitude and frequency,a live cell exhibits mechanical anisotropy and responds to a local shear stress differently from responding to a local complex stress by stretching chromatin and upregulating gene transcription to different levels,extending our previous finding on force-induced direct gene activation.This finding highlights the importance of force modes in impacting cellular mechanical and biological responses in living cells and tissues and may have implications in tissue patterning and embryonic development.展开更多
The technology of induced pluripotent stem cell(iPSCs)has enabled the conversion of somatic cells into primitive undifferentiated cells via reprogramming.This approach provides possibilities for cell replacement thera...The technology of induced pluripotent stem cell(iPSCs)has enabled the conversion of somatic cells into primitive undifferentiated cells via reprogramming.This approach provides possibilities for cell replacement therapies and drug screening,but the potential risk of tumorigenesis hampers further development and application.How to generate required differentia-ted cells without initiating tumor progression remains a huge challenge.Here we show that mouse embryonic fibroblasts could be differentiated into valvular endothelial cell(VEC)like cells.VECs are critical in valve replacements in aortic valve failure.VEC-associated gene and protein expression and functional assays were quantified for these VEC-like cells.We show that mouse embryonic fibroblasts could be converted into VEC-like cells.Our results suggest that it is possible to convert mouse embryonic fibroblasts into VEC-like cells without first reprogramming them into pluripotent stem cells,minimizing the possibility of tumorigenesis.展开更多
Recently we have synthesized a novel small retinoid molecule WYC-209 that can effectively inhibit proliferation of malignant murine melanoma tumor-repopulating cells(TRCs).The molecule can induce 100%TRCs apoptosis at...Recently we have synthesized a novel small retinoid molecule WYC-209 that can effectively inhibit proliferation of malignant murine melanoma tumor-repopulating cells(TRCs).The molecule can induce 100%TRCs apoptosis at 10μM concentration.However,how WYC-209 induces TRCs apoptosis is still elusive.Here we demonstrate that WYC-209 at>6μM concentration started to induce TRCs apoptosis primarily via the caspase 3 pathway by releasing cytochrome c from mitochondria.Interestingly,we found that at concentrations<6μM WYC-209 induced TRCs to elevate dormancy marker COUP TF1 but induced no changes in apoptosis marker P53.Furthermore,proliferation markers Ki67 and PCNA decreased with the increase of WYC-209 concentrations,suggesting that low concentrations of WYC-209 inhibit TRCs growth by inducing cell dormancy instead of causing apoptosis.In addition,TRC traction forces were almost abolished when WYC-209 concentration was at 5μM,preceding the initiation of apoptosis.Our findings demonstrate that inhibition of TRCs by anti-cancer molecule WYC-209 is concentration-dependent and WYC-209 inhibits cellular force generation of the tumor-repopulating cells before inducing apoptosis.展开更多
It is known that mechanical forces play critical roles in physiology and diseases but the underlying mechanisms remain largely unknown[1].Most studies on the role of forces focus on cell surface molecules and cytoplas...It is known that mechanical forces play critical roles in physiology and diseases but the underlying mechanisms remain largely unknown[1].Most studies on the role of forces focus on cell surface molecules and cytoplasmic proteins.However,increasing evidence suggests that nuclear mechanotransduction impacts nuclear activities and functions.Recently we have revealed that transgene dihydrofolate reductase(DHFR)gene expression is directly upregulated via cell surface forceinduced stretching of chromatin [2].Here we show that endogenous genes are also upregulated directly by force via integrins.We present evidence on an underlying mechanism of how gene transcription is regulated by force.We have developed a technique of elastic round microgels to quantify 3D tractions in vitro and in vivo[3].We report a synthetic small molecule(which has been stiffened structurally)that inhibits malignant tumor repopulating cell growth in a low-stiffness(force)microenvironment and cancer metastasis in mouse models without detectable toxicity[4].These findings suggest that direct nuclear mechanotransduction impacts mechanobiology and mechanomedicine at cellular and molecular levels.展开更多
Spinal cord injury repair is one of the major challenges in medicine,as it can lead to permanent loss of function of central nervous system and damage to other function of the body.Stem cell transplantation together w...Spinal cord injury repair is one of the major challenges in medicine,as it can lead to permanent loss of function of central nervous system and damage to other function of the body.Stem cell transplantation together with tissue engineering is increasingly becoming a potential choice of treatment.However,direct transplantation of stem cells without scaffolds has yielded poor clinical outcome.Here we show a strategy of using mouse embryonic stem cells(ESCs)cultured within a silk fibroin(SF)based,three-dimensional scaffold with oriented channels by a directional temperature field freezing technique and lysophilization.We find that the ESCs maintained proliferation and migrated in the scaffolds and the cells migrated fastest along the SF channels.SF scaffolds contributed to ESC differentiation into neural and glial cell like cells and expressions of the neural and glial cell markers MAP2 and GFAP were greatly elevated when retinoic acid was used as an inducing factor.Our results suggest that this approach may offer some hope in the future for spinal cord injury repair using SF scaffolds and ESCs.展开更多
Daidzein is always added into poultry feed to make the production performance and immunity of poultry better. In this study, a total of 600 40-week-old Hy-Line variety brown layers were randomized into five groups and...Daidzein is always added into poultry feed to make the production performance and immunity of poultry better. In this study, a total of 600 40-week-old Hy-Line variety brown layers were randomized into five groups and fed with a corn-soybean-mixed basal diet supplement with 0, 10, 50, 100, and 500 mg · kg^-1 daidzein, respectively. Then, two PCR-based typing methods（RAPD-PCR and ERIC-PCR） were combined to analyze the ileum content and explore the changes of ileum microflora biodiversity. The results of RAPD-PCR and ERIC-PCR showed that bands under 10 mg · kg^-1 and 50 mg · kg^-1 were the most, and their similarity was the largest. Bands under 500 mg · kg^-1 were the least and similarity with other groups was the minimum. Ileum microflora biodiversity under 10 mg · kg^-1 or 50 mg · kg^-1 was richer than that under 500 mg · kg^-1. A corn-soybean-mixed basal diet supplement with 10 mg · kg^-1 to 50 mg · kg^-1 of daidzein might be beneficial to Hy-Line variety brown layers intestinal bacteria.展开更多
Yersinia enterocolitica is an important zoonotic pathogen that can induce disease outbreaks in a wide host range. Strain YER6022 was isolated from Pelteobagrus vachelli and identified using bacterial morphology and 16...Yersinia enterocolitica is an important zoonotic pathogen that can induce disease outbreaks in a wide host range. Strain YER6022 was isolated from Pelteobagrus vachelli and identified using bacterial morphology and 16S rDNA sequence analysis. Five virulence factors were detected, then artificial infection experiment and histopathological method were carried out. These results showed that strain YER6022 was one of Y. enterocolitica family members. In addition, ail, ystb, virF, yadA and HPIint were dectected. In artificial infection experiment, with 80% mortality and 100% morbidity, injected Pelteobagrus vachellis showed red swollen of the anus, abdomen swelling and fim bleeding. There existed serious hyperaemia and edema in kidney, spleen, intestine and liver at the light microscope. Ultrastructural observation indicated that mitochondria of the liver, kidney, spleen and intestine swelled and mitochondrial cristae broke. The data had further shed light on its pathogenicity in Pelteobagrus vachelli. It would benefit for further studies on pathogenesis ofPelteobagrus vachelli infected with Y. enterocolitica.展开更多
Objective Hepatocyte nuclear factor 4-alpha(HNF4A)is a critical transcription factor in the liver and pancreas.Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1(MODY1).Notably,MODY1 patients with HN...Objective Hepatocyte nuclear factor 4-alpha(HNF4A)is a critical transcription factor in the liver and pancreas.Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1(MODY1).Notably,MODY1 patients with HNF4A pathogenic mutations exhibit decreased responses to arginine and reduced plasma triglyceride levels,but the mechanisms remain unclear.This study aims to investigate the potential target genes transcriptionally regulated by HNF4A and explore its role in these metabolic pathways.Methods A stable 293T cell line expressing the HNF1A reporter was overexpressed with HNF4A.RNA sequencing(RNA-seq)was performed to analyze transcriptional differences.Transcription factor binding site prediction was then conducted to identify HNF4A binding motifs in the promoter regions of relevant target genes.Results RNA-seq results revealed a significant upregulation of transmembrane 4 L six family member 5(TM4SF5)mRNA in HNF4A-overexpressing cells.Transcription factor binding predictions suggested the presence of five potential HNF4A binding motifs in the TM4SF5 promoter.Finally,we confirmed that the DR1 site in the-57 to-48 region of the TM4SF5 promoter is the key binding motif for HNF4A.Conclusion This study identified TM4SF5 as a target gene of HNF4A and determined the key binding motif involved in its regulation.Given the role of TM4SF5 as an arginine sensor in mTOR signaling activation and triglyceride secretion,which closely aligns with phenotypes observed in MODY1 patients,our findings provide novel insights into the possible mechanisms by which HNF4A regulates triglyceride secretion in the liver and arginine-stimulated insulin secretion in the pancreas.展开更多
All living cells in a human body are made of the same DNA molecule but cells in different tissues express different genes and proteins.How the transcription process is controlled and regulated is largely unknown.Speci...All living cells in a human body are made of the same DNA molecule but cells in different tissues express different genes and proteins.How the transcription process is controlled and regulated is largely unknown.Specifically,mechanical forces are increasingly recognized to play critical roles in cell and tissue functions.However,what controls force-induced gene transcription is elusive.Recently we have reported that a local surface force transfers from integrins to the cytoskeleton and the link of nucleoskeleton and the cytoskeleton(LINC)into the nucleus and deforms chromatin directly to induce rapid activation of transgene DHFR.Here we show that endogenous mechanoresponsive genes egr-1 and Cav1 are rapidly upregulated and their upregulation depends on stress angles relative to the cell long axis,suggesting direct impact of these genes by force.Demethylation of histone 3 at lysine 9(H3K9)trimethylation(H3K9me3)at nuclear interiors(euchromatin)is necessary for force-induced transcription upregulation.Our findings suggest that force-rapid upregulation of mechanoresponsive genes by force depends on H3K9me3 demethylation.展开更多
OBJECTIVE To investigate the clinical and functional association of mi R-194 in human laryngeal squamous cell carcinoma(LSCC).METHODS Cell growth was measured by MTT assay.Cel cycle distribution was detected using PI ...OBJECTIVE To investigate the clinical and functional association of mi R-194 in human laryngeal squamous cell carcinoma(LSCC).METHODS Cell growth was measured by MTT assay.Cel cycle distribution was detected using PI staining by flow cytometric analysis.Cell migration and invasion were examined by wound healing assay and transwell assay.The 3′-UTR activity was detected by luciferase assay.The expression level of proteins and mR NA were analysed by Western blotting,immunohistochemistry and q RT-PCR.Mouse xenograft model was established to observe the tumor growth in vivo.RESULTS The expression level of miR-194 is significantly lower in clinical LSCC tissues compared with normal tissues,and is correlated with lymph node metastasis and TNM stage.Kaplan-Meier analysis shows that high miR-194 expression predicts a favorable outcome for LSCC patients.Functional assays show that enforced expression of mi R-194 inhibits the growth,migration,invasion and drug-resistance of LSCC cells.Moreover,Wee1 is identified as a novel functional target of mi R-194.Exogenous expression of Wee1 protein in mi R-194-over expressing cells partially reverses the suppressive effects of mi R-194 on LSCC cells.In addition,Wee1 was abnormally overexpressed in clinical LSCC tissues,and its protein levels were inversely correlated with miR-194 expression.High Wee1 protein level was also associated with TNM stage,lymph node metastasis and poor prognosis.CONCLUSION Our study provides new sights into the role of miR-194/Wee1 axis in LSCC,and suggests a novel miR-194/Wee1-based clinical intervention target for LSCC patients.展开更多
基金Supported by the National Science Foundation of China(31771692)Major Project on Breeding of New Varieties of Genetically Modified Organisms(2011ZX08004-002)
文摘Saline-alkaline stress can dramatically inhibit plant growth and limit crop production. Wild soybean （Glycine soja） is a crop that adapts well to such environmental stresses. In this study, RNA-sequencing technology was used to analyze the transcriptome profles of G. soja roots subjected to 50 mmol·L^-1 NaHCO3 and 150 mmol·L^-1 NaCl treatments. Totally, 2 125 differentially-expressed genes （DEGs） after NaCl treatment and 1 839 DEGs after NaHCO3 treatment were identifed. The top 14 DEGs revealed by RNA-seq were analyzed using qRT-PCR （quantitative real-time polymerase chain reaction）. Gene ontology （GO） annotation showed that most of DEGs under salt and alkali stresses were enriched in ＂metabolic process＂, ＂catalytic activity＂ and ＂binding＂ terms. To search for transcription factors （TFs） among DEGs, the data were screened against TF database PlantTFDB, and it was found that fve TF families, Apetala2/ethylene-responsive element binding proteins （AP2-EREBP）, V-myb avian myeloblastosis viral oncogene homolog （MYB）, WRKYGQK and Zinc fnger motif （WRKY）, NAM, ATAF1/2, CUC1/2 （NAC） and Cys2/His2 （C2H2） were involved in salt stress response. Other fve TF families, NAC, WRKY, MYB, AP2-EREBP and bZIP were involved in response to alkali stress. These two stress treatments shared NAC, WRKY, AP2-EREBP and MYB, and the only two different TFs were bZIP and C2H2. Forty-eight MYB TFs were differentially expressed under salt and alkali stresses, and most of them were up-regulated. This study provided useful information for further investigation of DEGs and TFs in response to saline and alkaline stresses and helped in understanding the molecular basis of the response of G. soja to saline and alkaline stresses.
基金Supported by the Science&Technology Department of Sichuan Province(2013FZ0014)the Construction Project of Postgraduate Academic Degree in Southwest University for Nationalities(2015XWD-S071007)
文摘The outer membrane protein, ompA, ofAeromonas veronii has a role in the virulence of the organism and is a potential candidate for vaccine development. In this study, ompA I ofAeromonas veronii strain WA106 was cloned and sequenced, then, it was expressed in Escherichia coli BL21. The nucleotide sequence of ompA I gene was 1 023 base pairs （GenBank Accession NO.KC748024）, which showed 100% homology with that of A. veronii （NO.AB290200.1）. This predicted protein was composed of 340 amino acid residues. Its molecular weight was 35.78 ku and isoelectric point was 5.18. The protein was a hydrophilic protein containing alpha helix and random coil with percentage of 35.0% and 49.7%, respectively. The tertiary structure, quaternary structure prediction showed that ompA I protein contained two peptide chains. SDS-PAGE showed that the actual value of the fusion protein was consistent with the expected result. It will facilitate further study of the role of ompA I protein.
基金Supported by the Natural Science Foundation of Heilongjiang Province(C2017025)。
文摘Myoblast differentiation is an essential process during skeletal muscle development.C2C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro.Dehydrogenase/reductase(SDR family)member 3(DHRS3)is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol.Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation.However,the effect of DHRS3 on mouse muscle cell differentiation was unclear.The objective of current study was to determine if DHRS3 affected muscle cell differentiation,and if DHRS3 was involved in muscle regeneration.Protein expression was determined by western blot and immunofluorescence analysis.The activation and inhibition of DHRS3 increased and decreased C2C12 myoblast differentiation respectively,which indicated that DHRS3 could affect C2C12 myoblast differentiation.DHRS3 expression was significantly changed during muscle regeneration,with the regeneration of muscle injury,the expression of DHRS3 tended to increase first and then decrease.It suggested that DHRS3 might be involved in muscle regeneration.In summary,this study confirmed the involvement of DHRS3 in C2C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development.
基金Supported by Young Doctor Fund of Gansu Education Department,China(2021QB-034)。
文摘In order to obtain pure enzyme with high activity,two amylase producing strains were isolated from soil samples,and named as the strains LZ-10 and LZ-11.According to morphologic observation,physiology and biochemistry experiments,16S rRNA and gyrB gene analysis,the strains LZ-10 and LZ-11 were identified as Bacillus subtilis.Adopted the method of ammonium sulfate,DEAE-52 anion purify enzyme,finally used polyacrylamide gel electrophoresis(SDS-PAGE)to detect molecular weight.The strain LZ-10 had an amylase activity of 123.3 U·mL^(-1),a purification factor of 6.8,a recovery rate of 69.5% and an optimal temperature of 50℃.The amylase activity of the strain LZ-11 was 59.91 U·mL^(-1),the purification factor was 4.5,the recovery rate was 60.5%,and the optimum temperature was 55℃.The commodity enzyme derived from Bacillus subtilis was 37.5 U·mL^(-1).The relative molecular weight of amylase activity from the two strains was 55 ku.Both thermal stability and pH stability were higher than those of commercialized amylase.
基金Supported by Project of Kunming University (YJL11014)
文摘According to our previous study, saprophytic fungi Botrytis cinerea contained 579 predicted secretary proteins. Among them, we found that 122 of these proteins contained the highly conserved pathogenic-related host-targeting-motif RxLx within 100 residues adjacent to the signal peptide cleavage site. According to PEDNAT and COG of the GenBank database, the functions of this motif containing proteins included metabolism modification and cell secretion. We blasted them in GenBank and found 47.54% had highly conserved homologues in other species, among them 74.1% had putative functional domains. This suggests these proteins are presumably ancient and vertically transmitted within the species. Many of these domains belonged to proteins which played roles in the pathogenic process of other kinds of pathogens and some had already been proved to be pathogenic secretary proteins of Botrytis cinerea. So we postulated that proteins contained host-targeting-motif RxLx were candidates participating in the pathogenesis of Botrytis cinerea.
基金supported by funds Huazhong University of Science and Technology,Wuhan,China and from US National Institutes of Health
文摘It is well established that living cells and tissues respond to mechanical forces such as flow-related shear stresses in blood or interstitial space and complex tractional stresses at cell-matrix contacts and cell-cell contacts.However,how different modes of forces impact mechanical and biological responses is elusive.Here we describe a strategy of using the three-dimensional magnetic twisting cytometry(3D MTC)technology to apply forces in any desired directions to the same living cell.We reveal that for a fixed stress amplitude and frequency,a live cell exhibits mechanical anisotropy and responds to a local shear stress differently from responding to a local complex stress by stretching chromatin and upregulating gene transcription to different levels,extending our previous finding on force-induced direct gene activation.This finding highlights the importance of force modes in impacting cellular mechanical and biological responses in living cells and tissues and may have implications in tissue patterning and embryonic development.
基金supported by funds from Huazhong University of Science and Technology
文摘The technology of induced pluripotent stem cell(iPSCs)has enabled the conversion of somatic cells into primitive undifferentiated cells via reprogramming.This approach provides possibilities for cell replacement therapies and drug screening,but the potential risk of tumorigenesis hampers further development and application.How to generate required differentia-ted cells without initiating tumor progression remains a huge challenge.Here we show that mouse embryonic fibroblasts could be differentiated into valvular endothelial cell(VEC)like cells.VECs are critical in valve replacements in aortic valve failure.VEC-associated gene and protein expression and functional assays were quantified for these VEC-like cells.We show that mouse embryonic fibroblasts could be converted into VEC-like cells.Our results suggest that it is possible to convert mouse embryonic fibroblasts into VEC-like cells without first reprogramming them into pluripotent stem cells,minimizing the possibility of tumorigenesis.
基金supported by funds from Huazhong University of Science and Technology
文摘Recently we have synthesized a novel small retinoid molecule WYC-209 that can effectively inhibit proliferation of malignant murine melanoma tumor-repopulating cells(TRCs).The molecule can induce 100%TRCs apoptosis at 10μM concentration.However,how WYC-209 induces TRCs apoptosis is still elusive.Here we demonstrate that WYC-209 at>6μM concentration started to induce TRCs apoptosis primarily via the caspase 3 pathway by releasing cytochrome c from mitochondria.Interestingly,we found that at concentrations<6μM WYC-209 induced TRCs to elevate dormancy marker COUP TF1 but induced no changes in apoptosis marker P53.Furthermore,proliferation markers Ki67 and PCNA decreased with the increase of WYC-209 concentrations,suggesting that low concentrations of WYC-209 inhibit TRCs growth by inducing cell dormancy instead of causing apoptosis.In addition,TRC traction forces were almost abolished when WYC-209 concentration was at 5μM,preceding the initiation of apoptosis.Our findings demonstrate that inhibition of TRCs by anti-cancer molecule WYC-209 is concentration-dependent and WYC-209 inhibits cellular force generation of the tumor-repopulating cells before inducing apoptosis.
基金supported by funds from National Institutes of Health,USA and Huazhong University of Science and Technology,Wuhan,Chinathe support from Hoeft Professorship at University of Illinois at Urbana-Champaign
文摘It is known that mechanical forces play critical roles in physiology and diseases but the underlying mechanisms remain largely unknown[1].Most studies on the role of forces focus on cell surface molecules and cytoplasmic proteins.However,increasing evidence suggests that nuclear mechanotransduction impacts nuclear activities and functions.Recently we have revealed that transgene dihydrofolate reductase(DHFR)gene expression is directly upregulated via cell surface forceinduced stretching of chromatin [2].Here we show that endogenous genes are also upregulated directly by force via integrins.We present evidence on an underlying mechanism of how gene transcription is regulated by force.We have developed a technique of elastic round microgels to quantify 3D tractions in vitro and in vivo[3].We report a synthetic small molecule(which has been stiffened structurally)that inhibits malignant tumor repopulating cell growth in a low-stiffness(force)microenvironment and cancer metastasis in mouse models without detectable toxicity[4].These findings suggest that direct nuclear mechanotransduction impacts mechanobiology and mechanomedicine at cellular and molecular levels.
基金supported by funds from Huazhong University of Science and Technology,Wuhan,China
文摘Spinal cord injury repair is one of the major challenges in medicine,as it can lead to permanent loss of function of central nervous system and damage to other function of the body.Stem cell transplantation together with tissue engineering is increasingly becoming a potential choice of treatment.However,direct transplantation of stem cells without scaffolds has yielded poor clinical outcome.Here we show a strategy of using mouse embryonic stem cells(ESCs)cultured within a silk fibroin(SF)based,three-dimensional scaffold with oriented channels by a directional temperature field freezing technique and lysophilization.We find that the ESCs maintained proliferation and migrated in the scaffolds and the cells migrated fastest along the SF channels.SF scaffolds contributed to ESC differentiation into neural and glial cell like cells and expressions of the neural and glial cell markers MAP2 and GFAP were greatly elevated when retinoic acid was used as an inducing factor.Our results suggest that this approach may offer some hope in the future for spinal cord injury repair using SF scaffolds and ESCs.
基金Supported by the Science&Technology Department of Sichuan Province(2013FZ0014)the Construction Project of Postgraduate Academic Degree in Southwest University for Nationalities(2014XWD-S071007)the National Training Plan of Excellent Agricultural and Forestal Talents(2014)
文摘Daidzein is always added into poultry feed to make the production performance and immunity of poultry better. In this study, a total of 600 40-week-old Hy-Line variety brown layers were randomized into five groups and fed with a corn-soybean-mixed basal diet supplement with 0, 10, 50, 100, and 500 mg · kg^-1 daidzein, respectively. Then, two PCR-based typing methods（RAPD-PCR and ERIC-PCR） were combined to analyze the ileum content and explore the changes of ileum microflora biodiversity. The results of RAPD-PCR and ERIC-PCR showed that bands under 10 mg · kg^-1 and 50 mg · kg^-1 were the most, and their similarity was the largest. Bands under 500 mg · kg^-1 were the least and similarity with other groups was the minimum. Ileum microflora biodiversity under 10 mg · kg^-1 or 50 mg · kg^-1 was richer than that under 500 mg · kg^-1. A corn-soybean-mixed basal diet supplement with 10 mg · kg^-1 to 50 mg · kg^-1 of daidzein might be beneficial to Hy-Line variety brown layers intestinal bacteria.
基金Supported by the Science&Technology Department of Sichuan Province(2013FZ0014)the Construction Project of the Postgraduate Academic Degree in Southwest University for Nationalities(2013XWD-S071007)
文摘Yersinia enterocolitica is an important zoonotic pathogen that can induce disease outbreaks in a wide host range. Strain YER6022 was isolated from Pelteobagrus vachelli and identified using bacterial morphology and 16S rDNA sequence analysis. Five virulence factors were detected, then artificial infection experiment and histopathological method were carried out. These results showed that strain YER6022 was one of Y. enterocolitica family members. In addition, ail, ystb, virF, yadA and HPIint were dectected. In artificial infection experiment, with 80% mortality and 100% morbidity, injected Pelteobagrus vachellis showed red swollen of the anus, abdomen swelling and fim bleeding. There existed serious hyperaemia and edema in kidney, spleen, intestine and liver at the light microscope. Ultrastructural observation indicated that mitochondria of the liver, kidney, spleen and intestine swelled and mitochondrial cristae broke. The data had further shed light on its pathogenicity in Pelteobagrus vachelli. It would benefit for further studies on pathogenesis ofPelteobagrus vachelli infected with Y. enterocolitica.
文摘Objective Hepatocyte nuclear factor 4-alpha(HNF4A)is a critical transcription factor in the liver and pancreas.Dysfunctions of HNF4A lead to maturity onset diabetes of the young 1(MODY1).Notably,MODY1 patients with HNF4A pathogenic mutations exhibit decreased responses to arginine and reduced plasma triglyceride levels,but the mechanisms remain unclear.This study aims to investigate the potential target genes transcriptionally regulated by HNF4A and explore its role in these metabolic pathways.Methods A stable 293T cell line expressing the HNF1A reporter was overexpressed with HNF4A.RNA sequencing(RNA-seq)was performed to analyze transcriptional differences.Transcription factor binding site prediction was then conducted to identify HNF4A binding motifs in the promoter regions of relevant target genes.Results RNA-seq results revealed a significant upregulation of transmembrane 4 L six family member 5(TM4SF5)mRNA in HNF4A-overexpressing cells.Transcription factor binding predictions suggested the presence of five potential HNF4A binding motifs in the TM4SF5 promoter.Finally,we confirmed that the DR1 site in the-57 to-48 region of the TM4SF5 promoter is the key binding motif for HNF4A.Conclusion This study identified TM4SF5 as a target gene of HNF4A and determined the key binding motif involved in its regulation.Given the role of TM4SF5 as an arginine sensor in mTOR signaling activation and triglyceride secretion,which closely aligns with phenotypes observed in MODY1 patients,our findings provide novel insights into the possible mechanisms by which HNF4A regulates triglyceride secretion in the liver and arginine-stimulated insulin secretion in the pancreas.
基金supported by the funds from Huazhong University of Science and Technology and US NIH grant GM 072744
文摘All living cells in a human body are made of the same DNA molecule but cells in different tissues express different genes and proteins.How the transcription process is controlled and regulated is largely unknown.Specifically,mechanical forces are increasingly recognized to play critical roles in cell and tissue functions.However,what controls force-induced gene transcription is elusive.Recently we have reported that a local surface force transfers from integrins to the cytoskeleton and the link of nucleoskeleton and the cytoskeleton(LINC)into the nucleus and deforms chromatin directly to induce rapid activation of transgene DHFR.Here we show that endogenous mechanoresponsive genes egr-1 and Cav1 are rapidly upregulated and their upregulation depends on stress angles relative to the cell long axis,suggesting direct impact of these genes by force.Demethylation of histone 3 at lysine 9(H3K9)trimethylation(H3K9me3)at nuclear interiors(euchromatin)is necessary for force-induced transcription upregulation.Our findings suggest that force-rapid upregulation of mechanoresponsive genes by force depends on H3K9me3 demethylation.
基金The project supported by National Natural Science Foundation of China(31271444,81201726)the Guangdong Natural Science Funds(2015TQ01R350,2014A030313057)the Guangdong Science and Technology Program(2016A050502027,2013B021800088)
文摘OBJECTIVE To investigate the clinical and functional association of mi R-194 in human laryngeal squamous cell carcinoma(LSCC).METHODS Cell growth was measured by MTT assay.Cel cycle distribution was detected using PI staining by flow cytometric analysis.Cell migration and invasion were examined by wound healing assay and transwell assay.The 3′-UTR activity was detected by luciferase assay.The expression level of proteins and mR NA were analysed by Western blotting,immunohistochemistry and q RT-PCR.Mouse xenograft model was established to observe the tumor growth in vivo.RESULTS The expression level of miR-194 is significantly lower in clinical LSCC tissues compared with normal tissues,and is correlated with lymph node metastasis and TNM stage.Kaplan-Meier analysis shows that high miR-194 expression predicts a favorable outcome for LSCC patients.Functional assays show that enforced expression of mi R-194 inhibits the growth,migration,invasion and drug-resistance of LSCC cells.Moreover,Wee1 is identified as a novel functional target of mi R-194.Exogenous expression of Wee1 protein in mi R-194-over expressing cells partially reverses the suppressive effects of mi R-194 on LSCC cells.In addition,Wee1 was abnormally overexpressed in clinical LSCC tissues,and its protein levels were inversely correlated with miR-194 expression.High Wee1 protein level was also associated with TNM stage,lymph node metastasis and poor prognosis.CONCLUSION Our study provides new sights into the role of miR-194/Wee1 axis in LSCC,and suggests a novel miR-194/Wee1-based clinical intervention target for LSCC patients.
