Objective To explore the effects of epithelial cell adhesion molecule(EPCM) expression downregulation on the proliferation,invasion and drug sensitivity of ECAM+/cluster of differentiation 44 (CD44) + colorectal cancer stem cells.Methods Colorectal cancer stem cells (ECAM) were obtained from human colorectal cancer cell line LoVo by the method of tumor microspheres.The specific markers of colorectal cancer stem cells were identified and studied.Lipofectamine 2000 liposome was used to complete the transfection,which was divided into three groups:blank group (ECAMhjgh/CD44 + LoVo),control group (transfection of common inhibitors,lipofectamine 2000-inhibitors) and experimental group (ECAM) according to different treatment methods.They were cultured in the same way,the three group of cells in logarithmic phase were treated with irinotecan (drug concentrations were 1,5,10,15,20,25,30,35,40,45,50 mg · L-1) and capecitabine (drug concentrations were 50,100,150,200,250,300,350,400,450,500,550,600mg· L-1).Then,they continued to be cultured.The expression levels of ECAM mRNA in three groups of ceils were detected by real-time fluorescence quantitative detection (RT-PCR).The thiazolyl blue tetrazolium bromide (MTI)assay was used to detect the proliferation and drug sensitivity of the three groups before and after treatment with irinotecan and capecitabine.Transwell cells were used to detect the invasion ability of three groups of cells.Results The percentage of ECAMhjgh/CD44 + colorectal cancer stem cell in LoVo cell line was 0.95％,85.78％ before and after enrichment,and the difference was significant (P ＜ 0.05).The expression of ECAM mRNA in blank group,control group and experimental group were 8.17 ±0.64,7.94 ±0.83,2.16 ±0.12.Compared with blank group,the difference had significantly in two groups (P ＜ 0.05,P ＜ 0.01).Compared with control group,the difference in experimental group had significantly in two groups (P ＜ 0.05).It indicated that model was succeed prepared.Invasive cell in the three groups were 79.22 ± 5.25,80.12 ± 4.89,31.23 ± 2.36.Compared with blank group,the difference had significantly in two groups (P ＜0.05,P ＜0.01).Compared with control group,the difference in experimental group had significantly(P ＜0.05).The IC50of irinotecan on colorectal cancer stem cells in the three groups were (20.25 ±4.35),(19.22 ±3.99),(10.24 ± 2.04) mg · L-1.The IC50of capecitabine on colorectal cancer stem cells in the three groups were (320.13 ± 23.65),(315.79 ± 21.03),(250.22 ± 15.45) mg · L-1.Compared with control group,the difference in experimental group had significantly(P ＜0.05).Conclusion Targeted down-regulation of ECAM can effectively inhibit the proliferation and invasion of colon cancer stem cells,and enhance their sensitivity to drugs at the same time.

Objective To evaluate the clinical efficacy and safety of amifostine on elderly acute leukemia patients receiving chemotherapy.Methods Fifty-eight patients with acute leukemia treated with chemo-therapy and amifostine were recruited in this study and then divided into two groups, 28 cases in elderly group (≥60 years) and 30 cases in control group (<60 years).All the patients were given amifostine 600 mg· m-2 through intravenous injection 15 to 30 minutes prior chemothe-rapy for 4 cycles.The data of the influence of amifostine on chemotherapy-induced adverse reactions as well as patients′blood pressure were compared in two groups.Results There was no statistical difference in incidence rates of chemotherapy -induced adverse reactions in two groups (P>0.05).After chemotherapy, there were 82 (80.4%) and 102 (80.3%) cases showing decreasing systolic blood pressure in elderly group and control group, respectively, and 71 ( 69.6%) and 83 ( 62.9%) cases showing decreasing diastolic blood pressure ( P >0.05).Conclusion The application of amifostine on elderly acute leukemia patients who has received chemotherapy is safe and could relieve chemotherapy-induced adverse reactions.

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1 and FUS mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis of motor neurons derived from SOD1 and corrected iPSCs revealed 899 aberrant transcripts. Our work may shed light on discovery of early biomarkers and pathways dysregulated in ALS, as well as provide a basis for novel therapeutic strategies to treat ALS.
Amyotrophic Lateral Sclerosis ; genetics ; metabolism ; therapy ; Cell Line ; Clustered Regularly Interspaced Short Palindromic Repeats ; Genetic Therapy ; Genome-Wide Association Study ; Humans ; Induced Pluripotent Stem Cells ; metabolism ; Mutation, Missense ; RNA-Binding Protein FUS ; genetics ; metabolism ; Superoxide Dismutase-1 ; genetics ; metabolism

Xeroderma pigmentosum (XP) is a group of genetic disorders caused by mutations of XP-associated genes, resulting in impairment of DNA repair. XP patients frequently exhibit neurological degeneration, but the underlying mechanism is unknown, in part due to lack of proper disease models. Here, we generated patient-specific induced pluripotent stem cells (iPSCs) harboring mutations in five different XP genes including XPA, XPB, XPC, XPG, and XPV. These iPSCs were further differentiated to neural cells, and their susceptibility to DNA damage stress was investigated. Mutation of XPA in either neural stem cells (NSCs) or neurons resulted in severe DNA damage repair defects, and these neural cells with mutant XPA were hyper-sensitive to DNA damage-induced apoptosis. Thus, XP-mutant neural cells represent valuable tools to clarify the molecular mechanisms of neurological abnormalities in the XP patients.
DNA Damage ; DNA Repair ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Humans ; Induced Pluripotent Stem Cells ; metabolism ; pathology ; Male ; Models, Biological ; Mutation ; Neural Stem Cells ; metabolism ; pathology ; Xeroderma Pigmentosum ; genetics ; metabolism ; pathology

Werner syndrome (WS) is a premature aging disorder that mainly affects tissues derived from mesoderm. We have recently developed a novel human WS model using WRN-deficient human mesenchymal stem cells (MSCs). This model recapitulates many phenotypic features of WS. Based on a screen of a number of chemicals, here we found that Vitamin C exerts most efficient rescue for many features in premature aging as shown in WRN-deficient MSCs, including cell growth arrest, increased reactive oxygen species levels, telomere attrition, excessive secretion of inflammatory factors, as well as disorganization of nuclear lamina and heterochromatin. Moreover, Vitamin C restores in vivo viability of MSCs in a mouse model. RNA sequencing analysis indicates that Vitamin C alters the expression of a series of genes involved in chromatin condensation, cell cycle regulation, DNA replication, and DNA damage repair pathways in WRN-deficient MSCs. Our results identify Vitamin C as a rejuvenating factor for WS MSCs, which holds the potential of being applied as a novel type of treatment of WS.
Animals ; Ascorbic Acid ; pharmacology ; Cell Cycle Checkpoints ; drug effects ; Cell Line ; Cellular Senescence ; drug effects ; DNA Damage ; DNA Repair ; drug effects ; DNA Replication ; drug effects ; Disease Models, Animal ; Heterochromatin ; metabolism ; pathology ; Humans ; Mesenchymal Stem Cells ; metabolism ; pathology ; Mice ; Nuclear Lamina ; metabolism ; pathology ; Reactive Oxygen Species ; metabolism ; Telomere Homeostasis ; drug effects ; Werner Syndrome ; drug therapy ; genetics ; metabolism