Objective:To investigate the effects of hepatic fatty acid transporter 5 (FATP5) gene expression on the levels of free long-chain fatty acid (LCFA) in mice with metabolic associated fatty liver disease (MAFLD).Methods:From June to December 2022, a prospective study was conducted with three experimental groups: wild-type (WT) group, FATP5 gene expression negative (FATP5 -) group, and human FATP5 gene expression positive (hFATP5 +) group, with 10 mice in each group. Each group of mice was fed a high-fat diet for 16 weeks to establish a model of MAFLD. Hepatic tissue changes were observed using hematoxylin-eosin staining. The liver mass and liver coefficient of the mice were measured. Total cholesterol (TC), triglycerides (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid (UA), and LCFA levels were determined using an automatic biochemical analyzer. Blood glucose (Glu) levels were measured using a blood glucose analyzer. The liver mass and liver coefficient, TC and TG levels, AST and ALT levels, Glu and UA levels, and LCFA levels were compared among the three groups. Results:In the WT group, there was significant inflammatory cell infiltration within the hepatocytes and a large amount of fat accumulation. In the FATP5 - group, the inflammatory cell infiltration in the hepatocytes was mild with slight fat accumulation. In the hFATP5 + group, the inflammatory cell infiltration in the hepatocytes was severe, with great fat accumulation. The liver mass [(1.27 ± 0.25) g], liver coefficient (2.38 ± 0.19), TC [(1.82 ± 0.26) mmol/L], TG [(0.93 ± 0.24) mmol/L], AST [(169.95 ± 37.73) U/L], ALT [(95.36 ± 21.49) U/L], Glu [(8.34 ± 1.52) mmol/L], and UA [(74.32 ± 15.52) μmol/L] in the FATP5 - group were all significantly lower than those in the WT group [(1.61 ± 0.23) g, (2.71 ± 0.20), (2.31 ± 0.28) mmol/L, (1.34 ± 0.21) mmol/L, (278.31 ± 43.24) U/L, (147.32 ± 28.81) U/L, (10.52 ± 1.24) mmol/L, (96.28 ± 17.43) μmol/L], while the LCFA level [(3.57 ± 0.48) mg/L] in the FATP5 - group was significantly higher than that in the WT group [(2.63 ± 0.56) mg/L] ( t = 3.17, 3.78, 4.06, 4.07, 5.97, 4.57, 3.51, 2.98, 4.03, all P < 0.05). In the hFATP5 + group, the liver mass [(1.92 ± 0.30) g], liver coefficient (2.95 ± 0.23), TC [(2.59 ± 0.24) mmol/L], TG [(1.76 ± 0.35) mmol/L], AST [(341.22 ± 48.98) U/L], ALT [(189.45 ± 17.97) U/L], Glu [(13.21 ± 1.98) mmol/L], and UA [(117.74 ± 18.38) μmol/L] were all significantly higher than those in the WT group, while the LCFA level [(3.57 ± 0.48) mg/L] in the FATP5 + group was significantly lower than that in the WT group ( t = 2.59, 2.49, 2.40, 3.25, 3.04, 3.92, 3.64, 2.68, 3.19, all P < 0.05). Conclusions:The absence of FATP5 in the liver can elevate blood levels of LCFA in mice with MAFLD, reduce food intake, and help alleviate the symptoms of MAFLD.

Objective:To investigate the effects of hepatic fatty acid transporter 5 (FATP5) gene expression on the levels of free long-chain fatty acid (LCFA) in mice with metabolic associated fatty liver disease (MAFLD).Methods:From June to December 2022, a prospective study was conducted with three experimental groups: wild-type (WT) group, FATP5 gene expression negative (FATP5 -) group, and human FATP5 gene expression positive (hFATP5 +) group, with 10 mice in each group. Each group of mice was fed a high-fat diet for 16 weeks to establish a model of MAFLD. Hepatic tissue changes were observed using hematoxylin-eosin staining. The liver mass and liver coefficient of the mice were measured. Total cholesterol (TC), triglycerides (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid (UA), and LCFA levels were determined using an automatic biochemical analyzer. Blood glucose (Glu) levels were measured using a blood glucose analyzer. The liver mass and liver coefficient, TC and TG levels, AST and ALT levels, Glu and UA levels, and LCFA levels were compared among the three groups. Results:In the WT group, there was significant inflammatory cell infiltration within the hepatocytes and a large amount of fat accumulation. In the FATP5 - group, the inflammatory cell infiltration in the hepatocytes was mild with slight fat accumulation. In the hFATP5 + group, the inflammatory cell infiltration in the hepatocytes was severe, with great fat accumulation. The liver mass [(1.27 ± 0.25) g], liver coefficient (2.38 ± 0.19), TC [(1.82 ± 0.26) mmol/L], TG [(0.93 ± 0.24) mmol/L], AST [(169.95 ± 37.73) U/L], ALT [(95.36 ± 21.49) U/L], Glu [(8.34 ± 1.52) mmol/L], and UA [(74.32 ± 15.52) μmol/L] in the FATP5 - group were all significantly lower than those in the WT group [(1.61 ± 0.23) g, (2.71 ± 0.20), (2.31 ± 0.28) mmol/L, (1.34 ± 0.21) mmol/L, (278.31 ± 43.24) U/L, (147.32 ± 28.81) U/L, (10.52 ± 1.24) mmol/L, (96.28 ± 17.43) μmol/L], while the LCFA level [(3.57 ± 0.48) mg/L] in the FATP5 - group was significantly higher than that in the WT group [(2.63 ± 0.56) mg/L] ( t = 3.17, 3.78, 4.06, 4.07, 5.97, 4.57, 3.51, 2.98, 4.03, all P < 0.05). In the hFATP5 + group, the liver mass [(1.92 ± 0.30) g], liver coefficient (2.95 ± 0.23), TC [(2.59 ± 0.24) mmol/L], TG [(1.76 ± 0.35) mmol/L], AST [(341.22 ± 48.98) U/L], ALT [(189.45 ± 17.97) U/L], Glu [(13.21 ± 1.98) mmol/L], and UA [(117.74 ± 18.38) μmol/L] were all significantly higher than those in the WT group, while the LCFA level [(3.57 ± 0.48) mg/L] in the FATP5 + group was significantly lower than that in the WT group ( t = 2.59, 2.49, 2.40, 3.25, 3.04, 3.92, 3.64, 2.68, 3.19, all P < 0.05). Conclusions:The absence of FATP5 in the liver can elevate blood levels of LCFA in mice with MAFLD, reduce food intake, and help alleviate the symptoms of MAFLD.

Objective To construct lentivirus containing CXCR4 gene and transfect MCF-7 cells , and obtain CXCR4 high-expressing MCF-7 cells. Methods CXCR4 gene was amplified by RT-PCR to construct CXCR4/pSin-EF2, which was transfected into HEK293T cells with psPAX2 and pMD2G vector for lentivirus packing. Packaged lentivirus was used to transfect human breast cancer cells MCF-7, with empty lentivirus as control. CXCR4 mRNA and protein expression levels were detected by RT-PCR and Western blot before and after transfection. And flow cytometry was used to detecte cell surface CXCR4 expression. Results The recombinant plasmid CXCR4/pSin-EF2 was constructed successfully,identified by double digestion and sequencing, and transfected into HEK293T cells to obtain high-titer lentivirus. RT-PCR and Western blot confirmed that the expression of CXCR4 in MCF-7 cells increased significantly after CXCR4 lentivirus transfection. Flow cytometry results showed that the CXCR4 positive rate increased from 26.78% to 99.29%, while there is no significant difference in CXCR4 expression between vector-transfected MCF-7 cells and non-transfected MCF-7 cells. Conclusion CXCR4 lentivirus and the breast cancer cell line with high and stable expression of CXCR4 (MCF-7CXCR4) were successfully constructed.