ABSTRACT
Objective To investigate the effects of transcription factor 2(TCF2) overexpression on insulin resistance in HepG2 cells. Methods HepG2 cells were treated with high concentration of insulin(1×10-8mol/L) for 24 hours to induce insulin resistance (IR). Cells were divided into four groups:control group,IR group,IR+vector group and IR+TCF2 overexpression group. RT-qPCR and Western blot were performed to detect the expres-sion of TCF2. Glucose consumption and glycogen synthesis were assayed by glucose oxidase method and anthrone method respectively. Cell viability was evaluated by MTT assay. The activities of hexokinase and pyruvate kinase were detected by colorimetry. The protein level of IRS-1 and GLUT4 was detected by Western blot.Results Com-pared with control group,the decreased glucose consumption was observed in IR group(P＜0.05),indicating that insulin-resistance model was established successfully. The mRNA and protein expression of TCF2 was remarkably down-regulated in IR group as compared with control group. Compared with IR group,overexpression of TCF2 sig-nificantly improved glucose consumption, liver glycogen content, and the activities of hexokinase and pyruvate kinase (P＜0.05). Moreover,TCF2 overexpression up-regulated the protein expression of IRS-1 and GLUT4 (P＜0.05).Conclusions TCF2 overexpression ameliorates insulin resistance of HepG2 cells.
ABSTRACT
To study the pathogens incidences in cord blood and the efficiency of different detective methods, 60 samples were drawn and reserved from collected and processed cord blood, respectively. The BACTEC 9050 system, improved Martin/thioglycollate broth (22 degrees C) and thioglycollate broth (35 degrees C) were employed to detected bacteria (including fungus) at the same time. Two hundred and six cord blood serum samples were used to detect the HBV DNA and HCV RNA by molecular biology technique, HBsAg, Anti-HBC, Anti-HCV, Anti-HCMV-IgM, HTLV-1, HTLV-2, HIV-1 and HIV-2 by ELISA and RBC agglutination test were used to detect the TPHA. Results showed that using BACTEC 9050 system, the incidence of bacteria and fungus was 3.33% and 0% respectively in collected cord blood; in processed cord blood, the rates increased to 6.67% and 1.67%, respectively. The sensitivity of BACTEC 9050 was higher than that of Martin/thioglycollate broth (22 degrees C/35 degrees C) culture. In 206 serum samples, the positive rate of HBV DNA was 5.8%, HCV RNA was 2.4%, HBsAg was 2.4%, HCMV-IgM was 1.89%, HCV was 2.4% and Anti-HBC was 29.4%. In those samples that Anti-HBC was positive, the positive rate of HBV DNA was 6.7%. It was concluded that the incidences of microbiological contamination in cord blood were high. The routine culture system would lead to false negative results of obligate anaerobes. It was necessary to replace the current culture system with improved system, such as BACTEC 9050 system. The molecular biology technique would make up for the default of ELISA.
Subject(s)
Humans , Bacteremia , Epidemiology , Blood Specimen Collection , Fetal Blood , Microbiology , Virology , Fungemia , Epidemiology , Polymerase Chain Reaction , Probability , Viremia , EpidemiologyABSTRACT
Breast cancer susceptibility gene 1(BRCA1) plays an important role in breast cancer development and progression. BRCA1 encodes a 1863-amino acid protein with two BRCA1 C-terminal (BRCT) domains at its C-terminus, BRCT1 and BRCT2. Many cancer-predisposing mutations are located in the BRCT domains, which have been shown to induce chromatin unfolding by use of an approach that allows visualization of large-scale chromatin structure through lac repressor/lac operator recognition. To map the important region of BRCT domain (amino acid residues 1642-1736), six deletion mutant constructs were made. The chromatin structure assay showed that amino acid residues 1691-1721 are involved in the induction of chromatin unfolding. To further localize the critical amino acid residues, ten alanine scanning mutant constructs were made. The chromatin structure assay demonstrated that the 1707IAGGK1711 region is critical for the chromatin unfolding activity. Based on the mapped important region, Blast analysis identified a novel homologous protein. Mapping of the BRCT1 domain may aid in the presymptomatic risk assessment and provide a valuable tool for further study on the BRCT1 structure and function.