AIM To determine whether cell division cycle(Cdc)42 is regulated by microRNA(miR)-15 a in the development of pediatric inflammatory bowel disease(IBD).METHODS We cultured 293 T cells, used plasmids and performed dual-...AIM To determine whether cell division cycle(Cdc)42 is regulated by microRNA(miR)-15 a in the development of pediatric inflammatory bowel disease(IBD).METHODS We cultured 293 T cells, used plasmids and performed dual-luciferase assay to determine whether Cdc42 is a miR-15 a target gene. We cultured Caco-2 cells, and stimulated them with tumor necrosis factor(TNF)-α. We then employed lentiviruses to alter the expression of miR-15 a and Cdc42. We performed quantitative reverse transcription polymerase chain reaction(qRTPCR) and immunofluorescence to determine whether Cdc42 is regulated by miR-15 a in Caco-2 cells. Finally, we collected ileocecal tissue by endoscopy from patients and performed qRT-PCR to examine the expression of miR-15 a and Cdc42 in pediatric IBD patients.RESULTS Target Scan and dual-luciferase assay revealed thatCdc42 was a miR-15 a target gene. MiR-15 a expression increased(P = 0.0038) and Cdc42 expression decreased(P = 0.0013) in cells stimulated with TNF-α, and the expression of the epithelial junction proteins zona occludens(ZO)-1(P < 0.05) and E-cadherin(P < 0.001) decreased. Cdc42 levels decreased in miR-15 a-mimic cells(P < 0.001) and increased in miR-15 a inhibitor cells(P < 0.05). ZO-1 and E-cadherin decreased in mi R-15 a-mimic cells(P < 0.001) but not in the miR-15 a inhibitor + TNF-α cells. In Lv-Cdc42 + TNF-α cells, ZO-1 and E-cadherin expression increased compared to the Lv-Cdc42-NC + TNF-α(P < 0.05) or miR-15 a-mimic cells(P < 0.05). Fifty-four pediatric IBD patients were included in this study, 21 in the control group, 19 in the Crohn's disease(CD) active(AC) group, seven in the CD remission(RE) group, and seven in the ulcerative colitis(UC) group. MiR-15 a increased and Cdc42 decreased in the CD AC group compared to the control group(P < 0.05). miR-15 a decreased and Cdc42 increased in the CD RE group compared to the CD AC group(P < 0.05). miR-15 a was positively correlated with the Pediatric Crohn's disease Activity Index(PCDAI)(P = 0.006), while Cdc42 was negatively correlated with PCDAI(P = 0.0008). Finally, miR-15 a expression negatively correlated with Cdc42 in pediatric IBD patients(P = 0.0045).CONCLUSION Mi R-15 a negatively regulates epithelial junctions through Cdc42 in Caco-2 cells and pediatric IBD patients.展开更多
目的:探讨核蛋白样转录因子核受体亚家族2组E成员1(nuclear receptor subfamily 2 group E member 1,NR2E1)对儿童神经母细胞瘤细胞株IMR-32生长、分裂、增殖的影响。方法:应用LipofectamineTM2000将构建的针对核蛋白样转录因子NR2E1的s...目的:探讨核蛋白样转录因子核受体亚家族2组E成员1(nuclear receptor subfamily 2 group E member 1,NR2E1)对儿童神经母细胞瘤细胞株IMR-32生长、分裂、增殖的影响。方法:应用LipofectamineTM2000将构建的针对核蛋白样转录因子NR2E1的shiRNA质粒载体转染神经母细胞瘤细胞株IMR32,并通过细胞计数法观察细胞生长抑制效应,采用细胞免疫荧光染色检测神经母细胞瘤细胞株IMR32细胞分裂蛋白的表达。结果:核蛋白样转录因子NR2E1的shiRNA质粒转染神经母细胞瘤细胞株IMR32 48 h后,该细胞株生长缓慢;相关细胞核分裂蛋白表达受到明显的抑制。结论:核蛋白样转录因子NR2E1的shiRNA干扰质粒转染神经母细胞瘤细胞株IMR32后,抑制了神经母细胞瘤细胞IMR32的分裂和增殖。展开更多
The main aim of the article is to investigate the irrational and transcendental properties of certain real numbers by means of the factorial series and the factorial number system. The difference between the factorial...The main aim of the article is to investigate the irrational and transcendental properties of certain real numbers by means of the factorial series and the factorial number system. The difference between the factorial series and the factorial system is that the factorial series does not set an upper bound at a given place after the radix point, while in the factorial system (i – 1) is the maximal possible value for r<sub>i</sub> after the radix point. I give an extended definition of periodic numbers, and show the relationship between periodic and irrational numbers. I prove the transcendence of e by means of the factorial series and the factorial number system.展开更多
Background:Considering the increase in the proportion of lung adenocarcinoma(LUAD)cases among all lung cancers and its considerable contribution to cancer-related deaths worldwide,we sought to identify novel oncogenes...Background:Considering the increase in the proportion of lung adenocarcinoma(LUAD)cases among all lung cancers and its considerable contribution to cancer-related deaths worldwide,we sought to identify novel oncogenes to provide potential targets and facilitate a better understanding of the malignant progression of LUAD.Methods:The results from the screening of transcriptome and survival analyses according to the integrated Gene Expression Omnibus(GEO)datasets and The Cancer Genome Atlas(TCGA)data were combined,and a promising risk biomarker called meiotic nuclear divisions 1(MND1)was selectively acquired.Cell viability assays and subcutaneous xenograftmodelswere used to validate the oncogenic role ofMND1 in LUADcell proliferation and tumor growth.Aseries of assays,including mass spectrometry,co-immunoprecipitation(Co-IP),and chromatin immunoprecipitation(ChIP),were performed to explore the underlying mechanism.Results:MND1 up-regulation was identified to be an independent risk factor for overall survival in LUAD patients evaluated by both tissue microarray staining and third party data analysis.In vivo and in vitro assays showed that MND1 promoted LUAD cell proliferation by regulating cell cycle.The results of the Co-IP,ChIP and dual-luciferase reporter assays validated that MND1 competitively bound to tumor suppressor Kruppel-like factor 6(KLF6),and thereby protecting E2F transcription factor 1(E2F1)from KLF6-induced transcriptional repression.Luciferase reporter and ChIP assays found that E2F1 activated MND1 transcription by binding to its promoter in a feedback manner.Conclusions:MND1,KLF6,and E2F1 form a positive feedback loop to regulate cell cycle and confer DDP resistance in LUAD.MND1 is crucial for malignant progression and may be a potential therapeutic target in LUAD patients.展开更多
基金Supported by the National Natural Science Foundation of Shanghai,No.201540068
文摘AIM To determine whether cell division cycle(Cdc)42 is regulated by microRNA(miR)-15 a in the development of pediatric inflammatory bowel disease(IBD).METHODS We cultured 293 T cells, used plasmids and performed dual-luciferase assay to determine whether Cdc42 is a miR-15 a target gene. We cultured Caco-2 cells, and stimulated them with tumor necrosis factor(TNF)-α. We then employed lentiviruses to alter the expression of miR-15 a and Cdc42. We performed quantitative reverse transcription polymerase chain reaction(qRTPCR) and immunofluorescence to determine whether Cdc42 is regulated by miR-15 a in Caco-2 cells. Finally, we collected ileocecal tissue by endoscopy from patients and performed qRT-PCR to examine the expression of miR-15 a and Cdc42 in pediatric IBD patients.RESULTS Target Scan and dual-luciferase assay revealed thatCdc42 was a miR-15 a target gene. MiR-15 a expression increased(P = 0.0038) and Cdc42 expression decreased(P = 0.0013) in cells stimulated with TNF-α, and the expression of the epithelial junction proteins zona occludens(ZO)-1(P < 0.05) and E-cadherin(P < 0.001) decreased. Cdc42 levels decreased in miR-15 a-mimic cells(P < 0.001) and increased in miR-15 a inhibitor cells(P < 0.05). ZO-1 and E-cadherin decreased in mi R-15 a-mimic cells(P < 0.001) but not in the miR-15 a inhibitor + TNF-α cells. In Lv-Cdc42 + TNF-α cells, ZO-1 and E-cadherin expression increased compared to the Lv-Cdc42-NC + TNF-α(P < 0.05) or miR-15 a-mimic cells(P < 0.05). Fifty-four pediatric IBD patients were included in this study, 21 in the control group, 19 in the Crohn's disease(CD) active(AC) group, seven in the CD remission(RE) group, and seven in the ulcerative colitis(UC) group. MiR-15 a increased and Cdc42 decreased in the CD AC group compared to the control group(P < 0.05). miR-15 a decreased and Cdc42 increased in the CD RE group compared to the CD AC group(P < 0.05). miR-15 a was positively correlated with the Pediatric Crohn's disease Activity Index(PCDAI)(P = 0.006), while Cdc42 was negatively correlated with PCDAI(P = 0.0008). Finally, miR-15 a expression negatively correlated with Cdc42 in pediatric IBD patients(P = 0.0045).CONCLUSION Mi R-15 a negatively regulates epithelial junctions through Cdc42 in Caco-2 cells and pediatric IBD patients.
文摘目的:探讨核蛋白样转录因子核受体亚家族2组E成员1(nuclear receptor subfamily 2 group E member 1,NR2E1)对儿童神经母细胞瘤细胞株IMR-32生长、分裂、增殖的影响。方法:应用LipofectamineTM2000将构建的针对核蛋白样转录因子NR2E1的shiRNA质粒载体转染神经母细胞瘤细胞株IMR32,并通过细胞计数法观察细胞生长抑制效应,采用细胞免疫荧光染色检测神经母细胞瘤细胞株IMR32细胞分裂蛋白的表达。结果:核蛋白样转录因子NR2E1的shiRNA质粒转染神经母细胞瘤细胞株IMR32 48 h后,该细胞株生长缓慢;相关细胞核分裂蛋白表达受到明显的抑制。结论:核蛋白样转录因子NR2E1的shiRNA干扰质粒转染神经母细胞瘤细胞株IMR32后,抑制了神经母细胞瘤细胞IMR32的分裂和增殖。
文摘The main aim of the article is to investigate the irrational and transcendental properties of certain real numbers by means of the factorial series and the factorial number system. The difference between the factorial series and the factorial system is that the factorial series does not set an upper bound at a given place after the radix point, while in the factorial system (i – 1) is the maximal possible value for r<sub>i</sub> after the radix point. I give an extended definition of periodic numbers, and show the relationship between periodic and irrational numbers. I prove the transcendence of e by means of the factorial series and the factorial number system.
基金Project of Jiangsu Provincial Medical Talent,Grant/Award Number:ZDRCA2016033China Postdoctoral Science Foundation,Grant/Award Number:2018M640465+2 种基金National Natural Science Foundation of China,Grant/Award Numbers:81672295,81702265,81802277,81872378Research Program of Jiangsu Health Department,Grant/Award Number:LGY2016025Social Development Project of Jiangsu Province,Grant/Award Number:BE2019758。
文摘Background:Considering the increase in the proportion of lung adenocarcinoma(LUAD)cases among all lung cancers and its considerable contribution to cancer-related deaths worldwide,we sought to identify novel oncogenes to provide potential targets and facilitate a better understanding of the malignant progression of LUAD.Methods:The results from the screening of transcriptome and survival analyses according to the integrated Gene Expression Omnibus(GEO)datasets and The Cancer Genome Atlas(TCGA)data were combined,and a promising risk biomarker called meiotic nuclear divisions 1(MND1)was selectively acquired.Cell viability assays and subcutaneous xenograftmodelswere used to validate the oncogenic role ofMND1 in LUADcell proliferation and tumor growth.Aseries of assays,including mass spectrometry,co-immunoprecipitation(Co-IP),and chromatin immunoprecipitation(ChIP),were performed to explore the underlying mechanism.Results:MND1 up-regulation was identified to be an independent risk factor for overall survival in LUAD patients evaluated by both tissue microarray staining and third party data analysis.In vivo and in vitro assays showed that MND1 promoted LUAD cell proliferation by regulating cell cycle.The results of the Co-IP,ChIP and dual-luciferase reporter assays validated that MND1 competitively bound to tumor suppressor Kruppel-like factor 6(KLF6),and thereby protecting E2F transcription factor 1(E2F1)from KLF6-induced transcriptional repression.Luciferase reporter and ChIP assays found that E2F1 activated MND1 transcription by binding to its promoter in a feedback manner.Conclusions:MND1,KLF6,and E2F1 form a positive feedback loop to regulate cell cycle and confer DDP resistance in LUAD.MND1 is crucial for malignant progression and may be a potential therapeutic target in LUAD patients.
