Objective: To study the expression level of TRF1 (telomeric repeat binding factor 1) protein in human acute leukemia and relationship between expression level of TRF1 protein and telomerase. Methods: A quantitative We...Objective: To study the expression level of TRF1 (telomeric repeat binding factor 1) protein in human acute leukemia and relationship between expression level of TRF1 protein and telomerase. Methods: A quantitative Western-Blot technique was developed using anti-TRF133-277 monoclonal antibody and GST-TRF1 purity protein as a standard to further determine the ex-pression level of TRF1 protein in total proteins extracted from clinical specimens. Results: Bone marrow tissues of 20 acute leukemia patients were studied, 11 healthy donors’ bone marrows were taken as a control. The expression level of TRF1 protein was significantly higher (P<0.01) in normal bone marrow ((2.217±0.462) μg/μl) than that of acute leukemia patients ((0.754±0.343) μg/μl). But there was no remarkable difference between ALL and ANLL patients ((0.618±0.285) μg/μl vs (0.845±0.359) μg/μl, P>0.05). After chemotherapy, TRF1 expression level of patients with complete remission elevated ((0.772±0.307) μg/μl vs (1.683±0.344) μg/μl, P<0.01), but lower than that of normal ((2.217±0.462) μg/μl, P<0.01). There was no significantly difference after chemotherapy ((0.726±0.411) μg/μl vs (0.895±0.339) μg/μl, P>0.05). TRF1 expression level of patients with complete remission is higher than that of patients without complete remission ((1.683±0.344) μg/μl vs (0.895±0.339) μg/μl, P<0.01). All samples were determined for telomerase activity. It was confirmed that the activity of telomerase in normal bone marrow was lower than that of acute leukemia patients ((0.125±0.078) μg/μl vs (0.765±0.284) μg/μl, P<0.01). There was no significant difference of expression level of TRF1 protein between ALL and ANLL patients ((0.897±0.290) μg/μl vs (0.677±0.268) μg/μl, P>0.05). After chemotherapy, telomerase activity of patients with complete remission decreased ((0.393±0.125) μg/μl), but was still higher than that of normal ((0.125±0.078) μg/μl, P<0.01). Conclusion: The expression level of TRF1 protein has corre-lativity to the activity of telomerase (P<0.001).展开更多
Aging, subjected to scientific scrutiny, is extensively defined as a time-dependent decline in functions that involves the majority of organisms. The time-dependent accretion of cellular lesions is generally a univers...Aging, subjected to scientific scrutiny, is extensively defined as a time-dependent decline in functions that involves the majority of organisms. The time-dependent accretion of cellular lesions is generally a universal trigger of aging, while mitochondrial dysfunction is a sign of aging. Dysfunctional mitochondria are identified and removed by mitophagy, a selective form of macroautophagy. Increased mitochondrial damage resulting from reduced biogenesis and clearance may promote the aging process. The primary purpose of this paper is to illustrate in detail the effects of mitophagy on aging and emphasize the associations between mitophagy and other signs of aging, including dietary restriction, telomere shortening, epigenetic alterations, and protein imbalance.The evidence regarding the effects of these elements on aging is still limited. And although the understanding of relationship between mitophagy and aging has been long-awaited, to analyze details of such a relationship remains the main challenge in aging studies.展开更多
Kluyveromyces marxianus is a food-safe yeast with great potential for producing heterologous proteins.Improving the yield in K.marxianus remains a challenge and incorporating large-scale functional modules poses a tec...Kluyveromyces marxianus is a food-safe yeast with great potential for producing heterologous proteins.Improving the yield in K.marxianus remains a challenge and incorporating large-scale functional modules poses a technical obstacle in engineering.To address these issues,linear and circular yeast artificial chromosomes of K.marxianus(KmYACs)were constructed and loaded with disulfide bond formation modules from Pichia pastoris or K.marxianus.These modules contained up to seven genes with a maximum size of 15 kb.KmYACs carried telomeres either from K.marxianus or Tetrahymena.KmYACs were transferred successfully into K.marxianus and stably propagated without affecting the normal growth of the host,regardless of the type of telomeres and configurations of KmYACs.KmYACs increased the overall expression levels of disulfide bond formation genes and significantly enhanced the yield of various heterologous proteins.In high-density fermentation,the use of KmYACs resulted in a glucoamylase yield of 16.8 g/l,the highest reported level to date in K.marxianus.Transcriptomic and metabolomic analysis of cells containing KmYACs suggested increased flavin adenine dinucleotide biosynthesis,enhanced flux entering the tricarboxylic acid cycle,and a preferred demand for lysine and arginine as features of cells overexpressing heterologous proteins.Consistently,supplementing lysine or arginine further improved the yield.Therefore,KmYAC provides a powerful platform for manipulating large modules with enormous potential for industrial applications and fundamental research.Transferring the disulfide bond formation module via YACs proves to be an efficient strategy for improving the yield of heterologous proteins,and this strategy may be applied to optimize other microbial cell factories.展开更多
文摘Objective: To study the expression level of TRF1 (telomeric repeat binding factor 1) protein in human acute leukemia and relationship between expression level of TRF1 protein and telomerase. Methods: A quantitative Western-Blot technique was developed using anti-TRF133-277 monoclonal antibody and GST-TRF1 purity protein as a standard to further determine the ex-pression level of TRF1 protein in total proteins extracted from clinical specimens. Results: Bone marrow tissues of 20 acute leukemia patients were studied, 11 healthy donors’ bone marrows were taken as a control. The expression level of TRF1 protein was significantly higher (P<0.01) in normal bone marrow ((2.217±0.462) μg/μl) than that of acute leukemia patients ((0.754±0.343) μg/μl). But there was no remarkable difference between ALL and ANLL patients ((0.618±0.285) μg/μl vs (0.845±0.359) μg/μl, P>0.05). After chemotherapy, TRF1 expression level of patients with complete remission elevated ((0.772±0.307) μg/μl vs (1.683±0.344) μg/μl, P<0.01), but lower than that of normal ((2.217±0.462) μg/μl, P<0.01). There was no significantly difference after chemotherapy ((0.726±0.411) μg/μl vs (0.895±0.339) μg/μl, P>0.05). TRF1 expression level of patients with complete remission is higher than that of patients without complete remission ((1.683±0.344) μg/μl vs (0.895±0.339) μg/μl, P<0.01). All samples were determined for telomerase activity. It was confirmed that the activity of telomerase in normal bone marrow was lower than that of acute leukemia patients ((0.125±0.078) μg/μl vs (0.765±0.284) μg/μl, P<0.01). There was no significant difference of expression level of TRF1 protein between ALL and ANLL patients ((0.897±0.290) μg/μl vs (0.677±0.268) μg/μl, P>0.05). After chemotherapy, telomerase activity of patients with complete remission decreased ((0.393±0.125) μg/μl), but was still higher than that of normal ((0.125±0.078) μg/μl, P<0.01). Conclusion: The expression level of TRF1 protein has corre-lativity to the activity of telomerase (P<0.001).
基金supported by the Construction Project of Capacity Improvement Plan for Chongqing Municipal Health Commission affiliated unit (2019NLTS001)-ZS03174the operating grant to Chongqing Key Laboratory of Neurodegenerative, and Diseases (Grant No. 1000013)Chongqing Talent Project (Grant No. 2000062)。
文摘Aging, subjected to scientific scrutiny, is extensively defined as a time-dependent decline in functions that involves the majority of organisms. The time-dependent accretion of cellular lesions is generally a universal trigger of aging, while mitochondrial dysfunction is a sign of aging. Dysfunctional mitochondria are identified and removed by mitophagy, a selective form of macroautophagy. Increased mitochondrial damage resulting from reduced biogenesis and clearance may promote the aging process. The primary purpose of this paper is to illustrate in detail the effects of mitophagy on aging and emphasize the associations between mitophagy and other signs of aging, including dietary restriction, telomere shortening, epigenetic alterations, and protein imbalance.The evidence regarding the effects of these elements on aging is still limited. And although the understanding of relationship between mitophagy and aging has been long-awaited, to analyze details of such a relationship remains the main challenge in aging studies.
基金supported by the National Key Research and Development Program of China(Nos.2021YFA0910601 and 2021YFC2100203)Shanghai Municipal Education Commission(2021-03-52)Science and Technology Research Program of Shanghai(19DZ2282100).
文摘Kluyveromyces marxianus is a food-safe yeast with great potential for producing heterologous proteins.Improving the yield in K.marxianus remains a challenge and incorporating large-scale functional modules poses a technical obstacle in engineering.To address these issues,linear and circular yeast artificial chromosomes of K.marxianus(KmYACs)were constructed and loaded with disulfide bond formation modules from Pichia pastoris or K.marxianus.These modules contained up to seven genes with a maximum size of 15 kb.KmYACs carried telomeres either from K.marxianus or Tetrahymena.KmYACs were transferred successfully into K.marxianus and stably propagated without affecting the normal growth of the host,regardless of the type of telomeres and configurations of KmYACs.KmYACs increased the overall expression levels of disulfide bond formation genes and significantly enhanced the yield of various heterologous proteins.In high-density fermentation,the use of KmYACs resulted in a glucoamylase yield of 16.8 g/l,the highest reported level to date in K.marxianus.Transcriptomic and metabolomic analysis of cells containing KmYACs suggested increased flavin adenine dinucleotide biosynthesis,enhanced flux entering the tricarboxylic acid cycle,and a preferred demand for lysine and arginine as features of cells overexpressing heterologous proteins.Consistently,supplementing lysine or arginine further improved the yield.Therefore,KmYAC provides a powerful platform for manipulating large modules with enormous potential for industrial applications and fundamental research.Transferring the disulfide bond formation module via YACs proves to be an efficient strategy for improving the yield of heterologous proteins,and this strategy may be applied to optimize other microbial cell factories.
