Journal: Frontiers in bioengineering and biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering.

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: FIGURE 3 | Biocompatibility of 5% composite SFC scaffolds (w/v). The viability staining and H&E staining of cells in both scaffolds (A). SEM images (B) showing the cell growth pattern on the surface of both scaffolds. IHC detection (C) of ALB and CYP3A4 expression of C3A cells in both scaffolds. Quantitative analysis of IHC staining (D) and functional gene expression of C3A cells in both scaffolds (E). (*p < 0.05).

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP2 (dilution, 1:50; Proteintech), and Alexa Fluor®647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Staining, Expressing, Immunohistochemistry, Functional Assay, Gene Expression

Journal: Frontiers in bioengineering and biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering.

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: FIGURE 7 | Functional gene and protein expression in different culture conditions using RT-qPCR (A) and IF staining (B). Higher functional gene expression was found in the 3D co-cultures than in the 2D cultures and 3D monocultures (A). iHepLPC-Heps in both groups expressed ALB, CYP3A4, and MRP2 after 14 days of culture (B). (*p < 0.05).

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP2 (dilution, 1:50; Proteintech), and Alexa Fluor®647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Functional Assay, Expressing, Quantitative RT-PCR, Staining, Gene Expression

Journal: Frontiers in bioengineering and biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering.

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: FIGURE 8 | Functional evaluation of iHepLPC-Heps on composite SFC scaffolds. Albumin secretion of different hepatic cultures was assayed using ELISA (A). Urea synthesis of iHepLPC-Heps cultured under different conditions (B). Induction of CYP3A4 (C1) and CYP1A2 (C2) expression in response to stimulation with omeprazole and rifampicin, assayed using RT-qPCR. DiI-LDL uptake and CDCFDA staining (D) of iHepLPC-Heps were examined in both groups. (*p < 0.05).

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP2 (dilution, 1:50; Proteintech), and Alexa Fluor®647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Functional Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Quantitative RT-PCR, Staining

Journal: Frontiers in bioengineering and biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering.

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: FIGURE 10 | IHC detection of ALB, CYP3A4, and MRP2 expression in both groups after transplantation. Protein expressions increased gradually over time. Compared with monocultures, the co-culture group showed upregulated expressions at each time point. Scale bar, 50 µm.

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP2 (dilution, 1:50; Proteintech), and Alexa Fluor®647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Expressing, Transplantation Assay, Co-Culture Assay

Journal: Biochemia Medica

Article Title: Common P-glycoprotein ( ABCB1 ) polymorphisms do not seem to be associated with the risk of rivaroxaban-related bleeding events: Preliminary data

doi: 10.11613/BM.2024.020703

Figure Lengend Snippet: Genotypes at the ABCG2 , CYP3A4 , CYP3A5 and CYP2J2 polymorphisms, genotype-predicted phenotypes and use of substrates, inhibitors and inducers

Article Snippet: We determined additional gene polymorphisms with possible effect on rivaroxaban pharmacokinetics, and considered them as covariates (confounders): the common loss-of-function ABCG2 polymorphism c.421C>A, rs2231142 (assay ID 4362691 C__15854163_70), CYP3A4 * 1B , rs2740574 (assay ID 4362691 C___1837671_50), and * 22; rs35599367 (assay ID 4351379 C__59013445_10) and CYP3A5 * 3 , rs776746 (assay ID C__26201809_30) polymorphisms needed to determine CYP3A4 /5 genotype-predicted phenotype, and CYP2J2 A>T (rs11572325, assay ID C__30760106_10) and * 7 (rs890293, assay ID 4362691 C___9581699_80) polymorphisms to determine CYP2J2 phenotype.

Techniques: Activity Assay, Variant Assay

Journal: Biochemia Medica

Article Title: Common P-glycoprotein ( ABCB1 ) polymorphisms do not seem to be associated with the risk of rivaroxaban-related bleeding events: Preliminary data

doi: 10.11613/BM.2024.020703

Figure Lengend Snippet: Prevalence of relevant comorbidities, comedication, and ABCG2 c.421C> A polymorphism, CYP3A4/5 and CYP2J2 phenotypes after covariate balancing

Article Snippet: We determined additional gene polymorphisms with possible effect on rivaroxaban pharmacokinetics, and considered them as covariates (confounders): the common loss-of-function ABCG2 polymorphism c.421C>A, rs2231142 (assay ID 4362691 C__15854163_70), CYP3A4 * 1B , rs2740574 (assay ID 4362691 C___1837671_50), and * 22; rs35599367 (assay ID 4351379 C__59013445_10) and CYP3A5 * 3 , rs776746 (assay ID C__26201809_30) polymorphisms needed to determine CYP3A4 /5 genotype-predicted phenotype, and CYP2J2 A>T (rs11572325, assay ID C__30760106_10) and * 7 (rs890293, assay ID 4362691 C___9581699_80) polymorphisms to determine CYP2J2 phenotype.

Techniques: Variant Assay, Activity Assay, Filtration

Journal: Acta Pharmaceutica Sinica. B

Article Title: Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression

doi: 10.1016/j.apsb.2014.08.003

Figure Lengend Snippet: CYP3A4 CDS-luciferase activities are reduced by individual or combined shRNAs in CHL cells. (A) CHL cells co-transfected with CYP3A4 CDS-luciferase reporter, pRL-TK and different shRNA expression plasmids showed 20%–30% lower luciferase activities, compared to cells transfected with the pS-NC plasmid. (B) CHL cells transfected with CYP3A4 CDS-luciferase reporter, pRL-TK and different combinations of shRNA expression plasmids showed 40%–50% lower luciferase activities, compared to cells transfected with the pS-NC plasmid. Firefly luciferase activity was normalized to corresponding Renilla luciferase activity, and the control group was set as 100%. * P <0.05; *** P <0.001 compared to the corresponding control ( n =3 in each group, which refers to the number of independent transfection samples in a representative experiment).

Article Snippet: Antibodies of rabbit anti-human CYP3A4 were provided by AVIVA Systems Biology (California, U.S.A.).

Techniques: Luciferase, Transfection, shRNA, Expressing, Plasmid Preparation, Activity Assay, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression

doi: 10.1016/j.apsb.2014.08.003

Figure Lengend Snippet: Combined shRNAs are effective in targeting CYP3A4 CDS in HEK293 cells. (A) HEK293 cells transfected with CYP3A4 CDS-luciferase reporter and three shRNA expression plasmids showed about 50% lower luciferase activities, compared to cells transfected with the pS-NC plasmid. *** P <0.001 compared to the control ( n =3 in each group, which refers to the number of independent transfection samples in a representative experiment). (B) Fluorescent microscopic pictures of CYP3A4 expression in HEK293 cells at 48 h after transient transfection with CYP3A4 CDS-luciferase reporter plasmid alone (CYP3A4), along with control plasmid (pS-NC) or three shRNA expression plasmids (S1+S2+S3).

Article Snippet: Antibodies of rabbit anti-human CYP3A4 were provided by AVIVA Systems Biology (California, U.S.A.).

Techniques: Transfection, Luciferase, shRNA, Expressing, Plasmid Preparation, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression

doi: 10.1016/j.apsb.2014.08.003

Figure Lengend Snippet: The mixture of three shRNA plasmids inhibits endogenous CYP3A4 mRNA expression in HepG2 cells. (A) CYP3A4 mRNA expression was elevated 2-fold in HepG2 cells by rifampicin (50 μmol/L for 48 h), as determined by RT-PCR analysis. (B) and (C) The shRNA expression plasmid mixture suppressed CYP3A4 mRNA levels in HepG2 cells. (D) The shRNA expression plasmid mixture did not change CYP3A5 mRNA expression in the HepG2 cells. Relative CYP3A4/5 mRNA expression level was normalized to corresponding GAPDH mRNA level, and the control group was set as 100%. *** P <0.001, compared to the vehicle control. n =3 in each group, which refers to the number of independent transfection samples in a representative experiment.

Article Snippet: Antibodies of rabbit anti-human CYP3A4 were provided by AVIVA Systems Biology (California, U.S.A.).

Techniques: shRNA, Expressing, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Control, Transfection

Journal: Acta Pharmaceutica Sinica. B

Article Title: Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression

doi: 10.1016/j.apsb.2014.08.003

Figure Lengend Snippet: Expression of endogenous CYP3A4 protein is suppressed by combined shRNAs in HepG2 cells. CYP3A4 protein expression (A) in HepG2 cells (treated with rifampicin) was suppressed by about 50% (B) after the transfection with the shRNA expression plasmid mixture, compared to pS-NC control plasmid. CYP3A4 protein expression level was normalized to corresponding GAPDH level in HepG2 cells, and control group was set as 100%. *** P <0.001, compared to the negative control ( n =3 in each group, which refers to the number of independent transfection samples in a representative experiment).

Article Snippet: Antibodies of rabbit anti-human CYP3A4 were provided by AVIVA Systems Biology (California, U.S.A.).

Techniques: Expressing, Transfection, shRNA, Plasmid Preparation, Control, Negative Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Three new shRNA expression vectors targeting the CYP3A4 coding sequence to inhibit its expression

doi: 10.1016/j.apsb.2014.08.003

Figure Lengend Snippet: CYP3A4 shRNAs alter the chemosensitivity of cells. Transfection of HepG2 cells with CYP3A4 shRNAs reduces the sensitivity to GA (15:1) or GA (17:1), compared to the cells transfected with control plasmid. Cytotoxicity was determined by the MTT assay. The inhibition rate relative to the control (0%) was calculated for each concentration of drug, and the IC 50 value was estimated by fitting the data to a Hill equation .

Article Snippet: Antibodies of rabbit anti-human CYP3A4 were provided by AVIVA Systems Biology (California, U.S.A.).

Techniques: Transfection, Control, Plasmid Preparation, MTT Assay, Inhibition, Concentration Assay