Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Hepatitis C Virus Increases Free Fatty Acids Absorption and Promotes its Replication Via Down-Regulating GADD45α Expression

doi: 10.12659/MSM.899591

Figure Lengend Snippet: Comparison of liver function index and lipid-related indicators in each group (mean ±SD).

Article Snippet: The HCV RNA polymerase chain reaction (PCR) commercial kit was from Daan Gene Co. Ltd., Guangzhou, China; FFA detection kits, HDL-C kit, and LDL-C kit were purchased from Sekisui Medical Technology (Japan).

Techniques: Comparison, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Hepatitis C Virus Increases Free Fatty Acids Absorption and Promotes its Replication Via Down-Regulating GADD45α Expression

doi: 10.12659/MSM.899591

Figure Lengend Snippet: Correlation analysis of HCV RNA virus load and levels of serum lipid-related indicators in patient groups.

Article Snippet: The HCV RNA polymerase chain reaction (PCR) commercial kit was from Daan Gene Co. Ltd., Guangzhou, China; FFA detection kits, HDL-C kit, and LDL-C kit were purchased from Sekisui Medical Technology (Japan).

Techniques: Virus

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Hepatitis C Virus Increases Free Fatty Acids Absorption and Promotes its Replication Via Down-Regulating GADD45α Expression

doi: 10.12659/MSM.899591

Figure Lengend Snippet: The role of FFA and HCV in GADD45α expression. ( A ) GADD45α protein expression and mRNA level were detected in tumor tissues from liver cancer patients coinciding with chronic HCV infection. Low, medium, and high mean different serum viral loads. ( B ) CCK-8 assay showing the effect of different concentrations of FFA (up to 2 mM) on cell viability of Huh7 cells. Cell viability was expressed as percentage of untreated cells. Huh7 cells (2×10 5 ) were cultured in a 24-well plate to 80%–85% confluence and then treated with different concentrations of FFA ( C ) or infected with HCV ( D ) for 72 hours. mRNA level of GADD45α was quantified by RT-PCR, and protein expression was detected by Western blotting. Results are shown as mean ±SD of three independent experiments.

Article Snippet: The HCV RNA polymerase chain reaction (PCR) commercial kit was from Daan Gene Co. Ltd., Guangzhou, China; FFA detection kits, HDL-C kit, and LDL-C kit were purchased from Sekisui Medical Technology (Japan).

Techniques: Expressing, Infection, CCK-8 Assay, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Hepatitis C Virus Increases Free Fatty Acids Absorption and Promotes its Replication Via Down-Regulating GADD45α Expression

doi: 10.12659/MSM.899591

Figure Lengend Snippet: FFA promotes HCV replication in Huh7 cells. Huh7 cells (2×10 5 ) were cultured in a 24-well plate to 80%–85% confluence and then co-cultured with different concentrations of FFA with or without HCV (MOI of 1.5) for 72 hours. ( A ) HCV RNA copies were quantified by RT-PCR, and NS3 protein level was detected by Western blotting. ( B ) Quantification of the GADD45α protein levels and mRNA levels in Huh7 cells. Results are shown as mean ± SD of three independent experiments.

Article Snippet: The HCV RNA polymerase chain reaction (PCR) commercial kit was from Daan Gene Co. Ltd., Guangzhou, China; FFA detection kits, HDL-C kit, and LDL-C kit were purchased from Sekisui Medical Technology (Japan).

Techniques: Cell Culture, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Hepatitis C Virus Increases Free Fatty Acids Absorption and Promotes its Replication Via Down-Regulating GADD45α Expression

doi: 10.12659/MSM.899591

Figure Lengend Snippet: GADD45α plays a vital role in regulating FFA-mediated HCV replication. Huh7 cells were transfected with GADD45α siRNA, a negative control siRNA (scramble), or GADD45α over-expressed plasmid. Forty-eight hours later, GADD45α expression was identified ( A ), and cells were then infected with HCV and simultaneously treated with FFA (0.8 mM). After 72 h, the relative HCV-NS3 protein and RNA expression were detected by Western blotting and RT-PCR ( A ); intracellular lipid accumulation was detected by Bodipy-493/503 staining and visualized after 400× amplification ( B ). Results are shown as mean ±SD of three independent experiments.

Article Snippet: The HCV RNA polymerase chain reaction (PCR) commercial kit was from Daan Gene Co. Ltd., Guangzhou, China; FFA detection kits, HDL-C kit, and LDL-C kit were purchased from Sekisui Medical Technology (Japan).

Techniques: Transfection, Negative Control, Plasmid Preparation, Expressing, Infection, RNA Expression, Western Blot, Reverse Transcription Polymerase Chain Reaction, Staining, Amplification