ABSTRACT
OBJECTIVE@#This study aims to establish an effective and stable periodontal ligament cell line stably expressing human telomerase reverse transcriptase (hTERT) gene by using the adenovirus method.@*METHODS@#Polymerase chain reaction (PCR) was used to amplify the full length of hTERT gene to construct recombinant adenovirus plasmid pAd-pshuttle-cmv-hTERT. Packaged adenovirus particles were used for infection of human periodontal ligament cells. The expression levels of hTERT and osteogenic genes, such as alkaline phosphatase, Runt-related transcription factor 2, bone sialoprotein, osteocalcin, osteopontin, and collagen Ⅰ mRNA, were detected by quantitative real-time PCR (qRT-PCR). The ability of osteogenic differentiation was observed by alizarin red staining, and the cell proliferation was determined by CCK-8.@*RESULTS@#Adenovirus particles containing the hTERT gene were successfully constructed and infected with periodontal ligament cells. The infected cells were similar to normal periodontal ligament cells. The qRT-PCR results showed that hTERT and osteogenesis-associated genes were highly expressed in the periodontal ligament cell lines constructed by adenoviruses. Alizarin red staining showed that the periodontal ligament cell line had strong osteogenic differentiation capability. CCK-8 showed that the periodontal ligament cell line had strong proliferation capability.@*CONCLUSIONS@#The human periodontal ligament cell line with high efficiency and stable expression of hTERT was established by the adenovirus method, thereby providing an ideal cell line for studying the mechanism of periodontal regeneration.
Subject(s)
Humans , Adenoviridae , Alkaline Phosphatase , Cell Differentiation , Cell Line , Cell Proliferation , Osteogenesis , Periodontal Ligament , TelomeraseABSTRACT
@#ObjectiveTo observe the effect of stimulating “Zhiyang” (DU 9) with electric acupuncture apparatus on contents of cAMP and cGMP in plasma of experimental acute myocardial ischemic rabbits.Methods40 rabbits were randomly divided into four groups: the normal control group (A), model group (B), model plus electric acupuncture on “Neiguan” (P 6) group (C) and model plus electric acupuncture on DU 9 group (D). The acute myocardial ischemic model was established by pituitrin (2 U/kg) injected into the vein of rabbit. Electrocardiogram (ECG) of Ⅱ leads was observed and contents of cAMP and cGMP in plasma were tested after electric acupuncture by radio immunoassay.ResultsThe cAMP contents of the group C and group D were lower than that of the group B significantly ( P<0.01), but not different from that of the group A ( P>0.05), and there was no difference between the group C and group D. The contents of cGMP were not different among four groups ( P>0.05).ConclusionPuncturing DU 9 can raise the myocardial endurance to the insufficiency of blood and oxygen. The effects of electric acupuncture on DU 9 and P 6 are the same.