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OBJECTIVE@#To study the correlation between DNA methylation patterns and gene expression in Down syndrome (DS).@*METHODS@#Induced pluripotent stem cells (iPSCs) derived from normal controls and DS patients were subjected to whole genome bisulfite sequencing and differentially methylated region (DMR) screening. Statistical analysis for chromosomal and gene element distribution were carried out for DMR. Gene ontology (GO) and enrichment-based cluster analysis were used to explore the molecular function of differentially expressed genes.@*RESULTS@#A total of 1569 DMR were identified in iPSCs derived from DS patients, for which the proportion of hypermethylation in promoter regions was significantly greater than that of the genebody. No DMR enrichment was noted on chromosome 21. Hypermethylation of the promoter and genebody was predicted to be inhibitory for gene expression. Functional clustering revealed the pathways related to neurodevelopmental, stem cell pluripotency and organ size regulation to be significantly correlated with differentially methylated genes.@*CONCLUSION@#Extensive and stochastic anomalies of genome-wide DNA methylation has been discovered in iPSCs derived from DS patients, for which the pattern and molecular regulation of methylation were significantly different from those of normal controls. Above findings suggested that DNA methylation pattern may play a vital role in both the pathogenesis of neurodevelopmental disorders and other phenotypic abnormalities during early embryonic development.
Subject(s)
Female , Humans , Pregnancy , DNA Methylation , Down Syndrome/genetics , Induced Pluripotent Stem Cells , Promoter Regions, Genetic , Whole Genome SequencingABSTRACT
Objective To investigate the correlation between the severity of alcoholic fatty liver disease and the amount of fat in the abdominal cavity and the serum inflammatory factor IL-18 and IL-8. Methods From October 2016 to October 2017,one hundred and twenty patients with AFLD in the First Hospital of Hebei Medical University were divided into light,medium,heavy groups according to the severity of fatty lesions by color Doppler Ultrasound. There were 40 mild patients,50 moderate patients and 30 severe patients. Forty healthy subjects were selected as controls. All the participants underwent CT scanning. The intra-abdominal fat area (VAT),abdominal subcutaneous fat area (SAT) and total abdominal fat area (TA) were measured. The liver function was measured by biochemical analyzer and enzyme-linked immunoassay (ELISA). (ELSIA) IL-18 was detected and IL-8 was detected by radioimmunoassay. Results The VAT of the healthy control group and the mild,medium and severe AFLD group were (70. 28±10. 19),(114. 38 ± 9. 97),(146. 73±10. 19),(163. 38±12. 69) cm2. The TA of the healthy control group and the mild, medium and severe AFLD group were ( 256. 72± 34. 56),( 332. 19 ± 33. 28),( 387. 49± 32. 28),( 478. 19 ±31. 02) cm2. The SAT of the healthy control group and the light,medium and severe AFLD group were (156. 23±28. 19),(203. 43±27. 12),(246. 19±26. 89),(271. 19 ±27. 94) cm2,respectively. Aspartate aminotransferase (AST) of the healthy control group and the mild,medium and severe AFLD group were (18. 50±1. 12),(23. 50±1. 21),(25. 50±1. 24),(29. 50± 1. 43) U/L. Alanine aminotransferase (ALT) of the healthy control group and the light, medium and severe AFLD group were ( 18. 50 ± 2. 14), ( 26. 50 ±2. 22),(35. 50±2. 34),(38. 50±2. 11) U/L. γ-glutamyltransferaseof the healthy control group and the light,medium and severe AFLD group were ( 16. 50 ± 2. 11), ( 32. 50 ± 2. 23), ( 47. 50 ± 2. 31), ( 48. 00 ±2. 43) U/L,respectively. Compared with the healthy control group,VAT,TA,SAT,AST,ALT andγ-GT in the light,medium and heavy AFLD group showed statistically significant differences ( P＜0. 05) . Compared with the mild AFLD group, VAT, TA, SAT, AST, ALT and γ-GT in the medium and heavy AFLD group showed statistically significant differences ( P＜0. 05) . Compared with the moderate AFLD group,the VAT, TA,SAT, AST, ALT, and γ-GT of the severe AFLD group showed statistically significant differences ( P＜0. 05). The data of the three AFLD groups showed that the concentration of all indicators were increasing as the severity of fat deepened. IL-18 of the healthy control group and the light,medium and severe AFLD group were (45. 67±4. 51),(52. 18±5. 09),(59. 87±4. 98),(64. 18±5. 12) ng/L; IL-8 of the healthy control group and the light, medium and severe AFLD group were ( 78. 92 ± 5. 07), ( 115. 62 ± 4. 89), ( 223. 76 ± 6. 78),(286. 42±7. 02) g/L. Compared with every group,IL-18 and IL-8 of light,medium and severe AFLD group showed statistically significant differences (F＝1035. 67,2. 93×105,P＜0. 001); compared with mild AFLD group,IL-18 and IL-8 of medium and heavy group showed statistically significant differences;compared with moderate AFLD group,IL-18 and IL-8 of severe group AFLD showed statistically significant differences ( P＜0. 001) . The levels of inflammatory factors IL-18 and IL-8 increased with the severity of steatosis. The severity of AFLD was significantly positively correlated with VAT,TA,SAT,IL-18 and IL-8 ( r 0. 415(P＜0. 001), 0. 435 ( P＜0. 001), 0. 512 ( P＜0. 001), 0. 274 ( P＜0. 001 ), 0. 689 ( P ＜0. 001). Conclusion Fat control is an important measure to prevent AFLD. IL-18 and IL-8 can reflect the severity of liver injury in AFLD and have important significance in judging prognosis.
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BACKGROUND:Umbilical cord mesenchymal stem cells are plentifully and conveniently obtained with a high proliferative ability, and have opened up a new way to treat patients with liver failure as they can differentiate into hepatocyte-like cells.OBJECTIVE:To observe the safety and efficacy in the treatment of chronic liver failure by transplanting umbilical cord mesenchymal stem cells.METHODS:Using parallel contrast method, 50 patients with chronic liver failure were divided into two groups, namely a stem cell group and a control group, containing 25 patients in each group. For the first group, transplantation of umbilical cord mesenchymal stem cells, (1.4-2.3)×106/kg, 100 mL, was given on the basis of medical comprehensive treatment,while for the second group only simple medical comprehensive treatment was given. The injection was done every 15 days, totally three times. Liver functions, prothrombin activity, Model for End-Stage Liver Disease (MELD) score, clinical symptoms, survival and side effects of the patients were observed before and 2, 4, 12 and 24 weeks after the treatment.RESULTS AND CONCLUSION:Compared with the control group, the albumin level and prothrombin activity were significantly increased in the stem cell group 12 and 24 weeks after treatment (P 0.05). Four weeks after treatment, clinical symptoms of the stem cell group improved significantly in comparison with the control group (P < 0.05).During the 24-week follow-up, the survival rate in the stem cell group was significantly higher than that in the control group (P < 0.05). Additionally, there were no adverse reactions and liver cancer associated with the stem cell therapy.Results show that it is safe and effective to treat patients with chronic liver failure through the transplantation of umbilical cord mesenchymal stem cells, and the cell transplantation can significantly improve patients' survival rate.
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Objective To observe the safety and clinic effect of umbilical blood stem cell transplantation for the patients with chronic liver failure (CLF).Methods 44 patients with CLF were included in the research and divided into two groups,22 in control group received internal medicine treatment,the other 22 in treatment group received umbilical blood stem cell transplantation in addition to internal medicine treatment.The biochemical index,MELD scores,clinical symptoms,survival situation and adverse reaction of the patients were observed within 2,4,12 and 24 weeks.Results Albumin and prothrombin activity of treatment group were higher than those of control group,the MELD scores of the treatment group was lower than that of control group,the survival rate was higher than the control group,and the difference is significant between the two groups (P ＜ 0.05).There was no significant difference between the two groups in terms of alanine aminotransferase and total bilirubin (P ＞ 0.05).After 4 weeks treatment,fatigue,inappetite,abdominal distention and ascitic fluid of the treatment group were better than that of control group,the difference was statistically significant (P ＜ 0.05).Besides,the patients of the both groups did not have any adverse reaction or hepatocellular carcinoma.Conclusion Umbilical blood stem cell transplantation is safe and effective for the patients with CLF and can improve the survival rate of the patients.
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Objective:To investigate the significance of metronomic therapy against Helicobacter pylori (HP) in the prevention of delayed emesis caused by chemotherapy of gastric cancer compared with the routine therapy. Methods:HP infection was confirmed by carbon 14 breath test in 69 patients. Combined chemotherapy was employed for the first time in the patients, who were divided into groups A and B. Metronomic therapy was administered to group A (n=33). Briefly, triplex medication against Helicobacter bacil i triplex was oral y ad-ministered:20 mg of omeprazole and 0.5 g of amoxicillin twice daily, with 200 mg of tinidazole once daily. Oral administration in group A was performed for 14 days from the start of chemotherapy. Simultaneously, 5-HT3 antagonists were applied. By contrast, group B (n=36) was treated with the oral triplex medication against Helicobacter bacilli:20 mg of omeprazole and 1 g of amoxicillin twice daily, with 400 mg of tinidazole once daily. Oral administration in group B was performed for 7 days from the beginning of chemotherapy with simultaneous application of 5-HT3 antagonists. Both groups were simultaneously treated with the 5-HT3 antagonist granisetron at 3 mg once daily during the administration of anti-HP therapy. HP infection was evaluated by immunohistochemistry before and after treatment. Results:The total effective rate for emesis in group A was 84.85%, which was significantly higher than that in group B (55.56%). Among the patients in group A, 15.15%demonstrated delayed emesis, compared with 44.44%of the patients in group B;the number of individuals was significantly lower in group A than in group B. The average number of chemotherapy cycles in group A was significantly higher than that in group B at 3.1 cycles;the difference between groups was statistically significant (P<0.05). In addition, the HP infection in group B was significantly lower than that in group A (P<0.05). Conclusion:Compared with one week of treatment with the conventional dose, two weeks of low-dose metronomic therapy against HP during chemotherapy can significantly reduce chemotherapy induced delayed emesis and can significantly reduce the degree of HP infection in patients with gastric cancer with HP infection.
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Objective: To clone human T cell receptor (TCR) ? chain gene and express its encoding protein by baculoviral expression system in insect cells. Methods: TCR ? chain cDNA was cloned from normal human peripheral blood mononuclear cells (PBMC) by RT-PCR and inserted into baculoviral transfer vector. This vector was co-transfected with baculovirus into insect sf 9 cells. The recombinant protein expressed was identified by SDS-PAGE and flow cytometry with mouse anti-human ? chain monoclonal antibody. Results: Human TCR ? chain cDNA cloned and inserted into baculoviral vector specifically expressed protein in the insect sf 9 cells, accounting for about 11% of the total protein yield in the supernatant of cell lysate. The molecular weight of the recombinant protein determined by SDS-PAGE was identical to what we anticipated. The insect cells transfected with recombinant baculovirus were demonstrated by intracellular labeling flow cytometry to express ? chain protein. Conclusion: Human T cell receptor ? chain gene was successfully cloned from human PBMC, and its encoding protein was highly expressed in insect cells. The TCR ? chain protein obtained by bioengineering technique is useful for in depth biological function study.