Heat shock transcription factor(HSF)are essential regulators of heat shock protein(HSP)gene expression in plants and algae,contributing to their resilience against biotic and abiotic stresses.However,the localization,...Heat shock transcription factor(HSF)are essential regulators of heat shock protein(HSP)gene expression in plants and algae,contributing to their resilience against biotic and abiotic stresses.However,the localization,structure,phylogenetic relationship,and characteristics of PtHSF genes in microalgae,especially in diatom Phaeodactylum tricornutum,remain largely unexplored.This study presents a comprehensive analysis of the PtHSF gene family in P.tricornutum.A genome-wide analysis identified 68 PtHSF genes,which were classified into two distinct subfamilies:traditional and untraditional.Motif and structure analyses revealed evidence of multiple duplication events within the PtHSF gene family.Expression profiling revealed diurnal patterns,with 34 genes being downregulated during the light period and upregulated during the dark period,while 19 genes exhibited the opposite pattern.These findings suggest that PtHSF genes may have specialized functions during the diurnal cycle and play a crucial role in maintaining cellular homeostasis in response to various stresses.Notably,PtHSF16,30,and 43 genes exhibited higher expression levels,suggesting their potential importance.This study provides a valuable foundation for future investigations into the specific functions of HSFs under different stress conditions and their regulatory mechanisms in P.tricornutum and other microalgae.展开更多
Objective:To study and analyze the expression level of homeobox A6(HOXA6)in patients with liver cancer and its correlation with prognosis.Methods:From January 2020 to July 2021,43 patients with liver cancer who underw...Objective:To study and analyze the expression level of homeobox A6(HOXA6)in patients with liver cancer and its correlation with prognosis.Methods:From January 2020 to July 2021,43 patients with liver cancer who underwent surgery without prior radiotherapy or chemotherapy were selected.Liver cancer tissues and adjacent tissues were collected,and the expression levels of HOXA6 mRNA and protein were measured using RT-qPCR and Western blot.The expression level of HOXA6 in tumor tissues(without radiotherapy and chemotherapy)was compared with that in adjacent tissues.Additionally,the prognosis and clinical characteristics of patients with low HOXA6 expression were compared to those with high HOXA6 expression,and the factors influencing high HOXA6 expression in liver cancer patients were analyzed.Results:HOXA6 mRNA and protein expression levels in tumor tissues were significantly higher than in adjacent tissues(P<0.05).There was no significant difference in the 1-year and 2-year survival rates between the high HOXA6 expression group and the low HOXA6 expression group(P>0.05).However,the 3-year survival rate was lower in the high HOXA6 expression group compared to the low HOXA6 expression group(P<0.05).There were no statistically significant differences between the two groups in terms of gender,age,tumor diameter,alpha-fetoprotein levels,or hepatitis B virus DNA levels(P>0.05).However,significant differences were found in the number of tumor lesions,degree of differentiation,and the proportion of tumor metastasis between the two groups(P<0.05).Multivariate logistic regression analysis revealed that the number of tumor lesions,degree of differentiation,and tumor metastasis were influencing factors for high HOXA6 expression in liver cancer patients(P<0.05).Conclusion:HOXA6 expression levels are abnormally elevated in liver cancer patients,and higher HOXA6 expression is associated with a worse 3-year survival rate.The factors influencing HOXA6 expression include the number of tumor lesions,degree of differentiation,and tumor metastasis.展开更多
[Objectives]This study was conducted to obtain a Chinese hamster ovary cell line that stably expresses recombinant human coagulation factor X(rhFX),and to induce efficient expression of the target gene with different ...[Objectives]This study was conducted to obtain a Chinese hamster ovary cell line that stably expresses recombinant human coagulation factor X(rhFX),and to induce efficient expression of the target gene with different concentrations of methotrexate(MTX).[Methods]PCR was performed to obtain the rhFX gene,and a recombinant expression plasmid pOptiVEC-rhFX was constructed and subjected to double restriction endonuclease digestion and sequencing identification.CHO-DG44(DHFR-)cells were transfected by the liposome method,and the target protein was purified by affinity chromatography and detected by SDS-PAGE electrophoresis and Western blot.A cell line with efficient and stable expression of the target gene was obtained by increasing the concentration of MTX to select positive clones.[Results]PCR yielded a 1509 bp rhFX sequence,and the results of double digestion and sequencing showed that the constructed pOptiVEC-rhFX plasmid was correct.After transfection of cells,MTX significantly increased protein expression.When MTX reached 1.0μmol/L,the expression efficiency of the target protein was(9±0.27)μg/ml.The purity of the target protein purified by affinity chromatography was 93%,which could be used for subsequent experiments.The expression efficiency of rhFX in eukaryotic mammalian cells was improved by increasing MTX concentration,and an affinity chromatography purification process for the target protein was preliminarily established.[Conclusions]The results of this study provide data support for the expression and purification of rhFX,and will lay a solid foundation for the development of drugs related to rhFX.展开更多
基金fundings of educational and scientific research projects for young and middle-aged teachers in Fujian Province(Grant number:2022JAT220693)Natural Science Foundation of Guangdong Province(Grant numbers:2024A1515013029 and 2022A1515012141).
文摘Heat shock transcription factor(HSF)are essential regulators of heat shock protein(HSP)gene expression in plants and algae,contributing to their resilience against biotic and abiotic stresses.However,the localization,structure,phylogenetic relationship,and characteristics of PtHSF genes in microalgae,especially in diatom Phaeodactylum tricornutum,remain largely unexplored.This study presents a comprehensive analysis of the PtHSF gene family in P.tricornutum.A genome-wide analysis identified 68 PtHSF genes,which were classified into two distinct subfamilies:traditional and untraditional.Motif and structure analyses revealed evidence of multiple duplication events within the PtHSF gene family.Expression profiling revealed diurnal patterns,with 34 genes being downregulated during the light period and upregulated during the dark period,while 19 genes exhibited the opposite pattern.These findings suggest that PtHSF genes may have specialized functions during the diurnal cycle and play a crucial role in maintaining cellular homeostasis in response to various stresses.Notably,PtHSF16,30,and 43 genes exhibited higher expression levels,suggesting their potential importance.This study provides a valuable foundation for future investigations into the specific functions of HSFs under different stress conditions and their regulatory mechanisms in P.tricornutum and other microalgae.
基金Medical Science Research Fund of Guangdong Province(Funding No.A2024264)。
文摘Objective:To study and analyze the expression level of homeobox A6(HOXA6)in patients with liver cancer and its correlation with prognosis.Methods:From January 2020 to July 2021,43 patients with liver cancer who underwent surgery without prior radiotherapy or chemotherapy were selected.Liver cancer tissues and adjacent tissues were collected,and the expression levels of HOXA6 mRNA and protein were measured using RT-qPCR and Western blot.The expression level of HOXA6 in tumor tissues(without radiotherapy and chemotherapy)was compared with that in adjacent tissues.Additionally,the prognosis and clinical characteristics of patients with low HOXA6 expression were compared to those with high HOXA6 expression,and the factors influencing high HOXA6 expression in liver cancer patients were analyzed.Results:HOXA6 mRNA and protein expression levels in tumor tissues were significantly higher than in adjacent tissues(P<0.05).There was no significant difference in the 1-year and 2-year survival rates between the high HOXA6 expression group and the low HOXA6 expression group(P>0.05).However,the 3-year survival rate was lower in the high HOXA6 expression group compared to the low HOXA6 expression group(P<0.05).There were no statistically significant differences between the two groups in terms of gender,age,tumor diameter,alpha-fetoprotein levels,or hepatitis B virus DNA levels(P>0.05).However,significant differences were found in the number of tumor lesions,degree of differentiation,and the proportion of tumor metastasis between the two groups(P<0.05).Multivariate logistic regression analysis revealed that the number of tumor lesions,degree of differentiation,and tumor metastasis were influencing factors for high HOXA6 expression in liver cancer patients(P<0.05).Conclusion:HOXA6 expression levels are abnormally elevated in liver cancer patients,and higher HOXA6 expression is associated with a worse 3-year survival rate.The factors influencing HOXA6 expression include the number of tumor lesions,degree of differentiation,and tumor metastasis.
基金Supported by Anhui Provincial Natural Science Foundation of China(2008085MC65)Natural Science Foundation of Anhui Higher Education Institutions of China(KJ2021A0922)+1 种基金China Postdoctoral Science Foundation(2020T130117ZX,2020M671914)Research Activities of Postdoctoral Researchers Foundation of Anhui Province,China(2020B470)。
文摘[Objectives]This study was conducted to obtain a Chinese hamster ovary cell line that stably expresses recombinant human coagulation factor X(rhFX),and to induce efficient expression of the target gene with different concentrations of methotrexate(MTX).[Methods]PCR was performed to obtain the rhFX gene,and a recombinant expression plasmid pOptiVEC-rhFX was constructed and subjected to double restriction endonuclease digestion and sequencing identification.CHO-DG44(DHFR-)cells were transfected by the liposome method,and the target protein was purified by affinity chromatography and detected by SDS-PAGE electrophoresis and Western blot.A cell line with efficient and stable expression of the target gene was obtained by increasing the concentration of MTX to select positive clones.[Results]PCR yielded a 1509 bp rhFX sequence,and the results of double digestion and sequencing showed that the constructed pOptiVEC-rhFX plasmid was correct.After transfection of cells,MTX significantly increased protein expression.When MTX reached 1.0μmol/L,the expression efficiency of the target protein was(9±0.27)μg/ml.The purity of the target protein purified by affinity chromatography was 93%,which could be used for subsequent experiments.The expression efficiency of rhFX in eukaryotic mammalian cells was improved by increasing MTX concentration,and an affinity chromatography purification process for the target protein was preliminarily established.[Conclusions]The results of this study provide data support for the expression and purification of rhFX,and will lay a solid foundation for the development of drugs related to rhFX.
