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Objective To construct lentivirus vectors carrying 16 short hairpin RNA (shRNA) expression cassettes targeting histone acetyltransferases and provide a powerful research approach to explore the mechanism of epigenetic genes in regulating hepatitis B virus (HBV).Methods Following the rule of short shRNA primer design,eight-pair primers (A ~ H )for each gene,which had stable interfering efficiency,were designed.The annealed primers were connected to the empty lentiviral vectors of shRNA for transformation.In order to confirm the positive clones,clones were analyzed by real-time polymerase chain reaction (RT-PCR ).Then, qualified plasmids were verified by enzyme digestion technology.Four shRNA lentivirus plasmids against the same gene were mixed to build lentivirus respectively.After the virus transfected into 293T cells for 48 and 72 hours,supernatants were collected to infect HepG2.2.15 cells.The percentage of fluorescent cells were observed and assessed by microscope 72 hours after infection.Results One hundred and twenty-eight lentiviral vectors of RNA interference (RNAi)library were constructed against 16 histone acetyltransferases and more than 80% of HepG2.2.15 cells were infected with lentivirus 72 hours after infection.Conclusions Sixteen shRNA lentivirus vectors against histone acetyltransferase are successfully constructed.Thus,a solid foundation for the study of the effect of human histone deacetylase on HBV replication is established.
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The diagnostic value of keratin-18 and its fragments in liver diseases has drawn increasingly interests in recent years,but studies on its application in chronic hepatitis B is still few.This paper reviews the circulating levels of keratin-18 fragments M30 and M65 in chronic hepatitis B and its application in the future.
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Objective To study the healing effect of recombinant human acidic fibroblast growth factor(rhaFGF) expressed in E.coli on dermal chronic ulcers in diabetic rats.Methods Ten male Wistar rats were used to set up diabetic dermal chronic ulcers models.The wounds were sprinkled with rhaFGF and physiological saline,respectively.The wound area,wound cavity volume and healing time were recorded,granulation tissue growth and epithelization in wound were observed,and wound healing status was evaluated.Results Compared with control group,the wound area and wound volume at different day in rhaFGF group were significantly diminuted(P
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0.05).Conclusion Among the DM patients,the incidence rate of PC is obviously increasing,and with the course of DM elongating,the risk of PC is increasing.There is no obvious difference between sexes.Compared with common people,the incidence rate of PC in DM patients is higher.Type 2 diabetes mellitus might be initial symptom of PC.
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Aim To investigate the effect of Panax Quinquefolium Saponin (PQS) on the cell immune system of the patients with Chronic Pulmonary Heart Disease (CPHD) and then to find out the relationship between the immune system and the mechanism of CPHD. Methods The T lymph subgroup and NK (CD3 -/CD16+56 +) cells in patients’ peripheral blood were detected by flow cytometry, and the expression of IL 2, IFN? mRNA were analyzed by RT PCR. Results CD3 +, CD4 + and CD4/CD8 in patients with CPHD were significantly lower than those in the control, while CD8 + higher ( P
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Objective:To investigate the association of thyrotropin receptor antibodies(TRAb) with the sensitivity to glucocorticoid therapy in Graves′s ophthalmopathy(GO) patients.Methods:Total 37 subjects with active GO were treated with prednisone. According to the relieve degree and clinical activity of their eye symptoms post-steroid treatment, patients were grouped into sensitive group(S) and non-sensitive group(NS). TRAb positive rate and antibody titer of the two groups were compared.Results:In S group( n=28 ), the positive rate and titre of TRAb were 76.53% and (12.54?5.62) U/L, respectively; The positive rate and titre of TRAb post-treatment were 22.18% and (7.82?4.91) U/L, respectively, were significant lower than data prior treatment. In NS group( n=9 ), the positive rate and titre of TRAb prior treatment were 67.24% and (11.07?4.63) U/L, respectively; The positive rate and titre of TRAb post-treatment were 59.74% and (10.81?5.96) U/L, there was no significant difference with data prior treatment. Prior treatment, there was no significant difference of TRAb titres between the two groups. Post-treatment, TRAb titres in NS group were significant higher than in S group.Conclusion:The variations of TRAb level were closely correlated with the sensitivity to glucocorticoid therapy in GO patients. Serum TRAb level can be used as a major index to evaluate therapeutic sensitivity to glucocorticoid therapy in GO.
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Objective:To observe the expression and correlation of eIF4E and c-myc in laryngeal squamous cell carcinoma.Methods:Western blot was applied to examine the expression of eIF4E and c-myc on tumor core,transition core and tumor-free core from 36 laryngeal carcinoma patients.The results were statistically analyzed.Results:The overexpression of eIF4E and c-myc protein levels suggests an increasing tendency from tumor-free core,transition core and tumor core;and shows an significant correlation of eIF4E and c-myc expression.Conclusion:The overexpression of eIF4E and c-myc cause the laryngeal carcinoma cell malignant transformation;the correlation of eIF4E and c-myc may provide a basis for gene therapy of laryngeal carcinoma.
