Objective To investigate the mechanism of polyethylene glycol losenatide in improving non-alcoholic fatty liver disease by regulating lipophagy.Methods(1)32 C57BL/6 male mice were fed normally for 1 week and randomly divided into 4 groups with 8 mice in each group.Normal control group(fed normal diet),Normal+GLP-1RA(intraperitoneal injection of polyethylene glycol losenatide 0.3 mg/kg q3d),High fat group(fed with high fat diet),High-fat+GLP-1RA(on the basis of high-fat diet,intraperitoneally injected polyethylene glycol losenatide 0.3 mg/kg q3d),The molding lasts 16 weeks.Body weight,blood glucose and glucose tolerance test(GTT)were measured at the end of 16 weeks.Serum total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),high density lipoprotein(HDL)and other indexes were detected by Enzyme-linked immunosor-bent assay(ELISA),liver pathological changes were observed by Hematoxylin-eosin(HE)and oil red O sections,and the expression of microtubule-associated protein light chain 3(LC3),autophagy adaptor protein(P62),and transcription factor EB(TFEB).in liver were detected by Western blot(Wb).(2)The cell was divided into normal group(treated with bovine serum albumin,BSA);Normal+polyethylene glycol losenatide group(BSA+100 nmol/L GLP-1RA),High fat group(0.5 mmol/L PA);High fat+polyethylene glycol losenatide group(0.5 mmol/L PA+100 nmol/L GLP-1RA).After 24 h,the steatosis was observed by oil red O staining,the levels of TG in the supernatant were detected,Wb analysis was performed to detect protein expression levels of LC3,TFEB,and P62.Reverse transcription quantitative PCR(RT-qPCR)was used to measure mRNA levels of lipid synthesis-related factors.Results(1)Compared with normal control group,the body weight and blood glucose of mice in the high fat group increased significantly(P<0.01),reduced glucose tolerance(P<0.01),the levels of TC,TG and LDL in serum in liver of high fat group were significantly increased(P<0.01),while HDL was significantly decreased(P<0.01).Compared with high fat group,blood sugar in mice,the levels of TC,TG and LDL in serum and P62 in liver of high fat+GLP-1 group were significantly decreased(P<0.01);Glucose tolerance,HDL and and the protein expressions of LC3,TFEB were significantly increased(P<0.01).Compared with normal control group,the above indexes in normal+GLP-1 group were not significantly changed(P>0.05).(2)In vitro,The NAFLD HepG2 cell model was successfully constructed with 0.5 mmol/L PA.After 24 h of PA treatment,intracellular lipid drops were significantly increased(P<0.01),intracellular TG and the mRNA levels of lipid synthesis-related factors in the cells were increased(P<0.01).High fat+GLP-1 group improved the fatty degeneration induced by high fat,and the changes of the above indexes showed an opposite trend but the expres-sion of LC3,TFEB were significantly increased(P<0.01),lipid regulatory factor P62 protein in liver were decreased(P<0.01).Conclusion Polyethylene glycol losenatide can improve non-alcoholic fatty liver disease by regulating lipophagy.

Objective To investigate the mechanism of polyethylene glycol losenatide in improving non-alcoholic fatty liver disease by regulating lipophagy.Methods(1)32 C57BL/6 male mice were fed normally for 1 week and randomly divided into 4 groups with 8 mice in each group.Normal control group(fed normal diet),Normal+GLP-1RA(intraperitoneal injection of polyethylene glycol losenatide 0.3 mg/kg q3d),High fat group(fed with high fat diet),High-fat+GLP-1RA(on the basis of high-fat diet,intraperitoneally injected polyethylene glycol losenatide 0.3 mg/kg q3d),The molding lasts 16 weeks.Body weight,blood glucose and glucose tolerance test(GTT)were measured at the end of 16 weeks.Serum total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),high density lipoprotein(HDL)and other indexes were detected by Enzyme-linked immunosor-bent assay(ELISA),liver pathological changes were observed by Hematoxylin-eosin(HE)and oil red O sections,and the expression of microtubule-associated protein light chain 3(LC3),autophagy adaptor protein(P62),and transcription factor EB(TFEB).in liver were detected by Western blot(Wb).(2)The cell was divided into normal group(treated with bovine serum albumin,BSA);Normal+polyethylene glycol losenatide group(BSA+100 nmol/L GLP-1RA),High fat group(0.5 mmol/L PA);High fat+polyethylene glycol losenatide group(0.5 mmol/L PA+100 nmol/L GLP-1RA).After 24 h,the steatosis was observed by oil red O staining,the levels of TG in the supernatant were detected,Wb analysis was performed to detect protein expression levels of LC3,TFEB,and P62.Reverse transcription quantitative PCR(RT-qPCR)was used to measure mRNA levels of lipid synthesis-related factors.Results(1)Compared with normal control group,the body weight and blood glucose of mice in the high fat group increased significantly(P<0.01),reduced glucose tolerance(P<0.01),the levels of TC,TG and LDL in serum in liver of high fat group were significantly increased(P<0.01),while HDL was significantly decreased(P<0.01).Compared with high fat group,blood sugar in mice,the levels of TC,TG and LDL in serum and P62 in liver of high fat+GLP-1 group were significantly decreased(P<0.01);Glucose tolerance,HDL and and the protein expressions of LC3,TFEB were significantly increased(P<0.01).Compared with normal control group,the above indexes in normal+GLP-1 group were not significantly changed(P>0.05).(2)In vitro,The NAFLD HepG2 cell model was successfully constructed with 0.5 mmol/L PA.After 24 h of PA treatment,intracellular lipid drops were significantly increased(P<0.01),intracellular TG and the mRNA levels of lipid synthesis-related factors in the cells were increased(P<0.01).High fat+GLP-1 group improved the fatty degeneration induced by high fat,and the changes of the above indexes showed an opposite trend but the expres-sion of LC3,TFEB were significantly increased(P<0.01),lipid regulatory factor P62 protein in liver were decreased(P<0.01).Conclusion Polyethylene glycol losenatide can improve non-alcoholic fatty liver disease by regulating lipophagy.

Objective:To assess the efficacy and prognosis of gastrointestinal stromal tumors(GIST)after preoperative targeted therapy us-ing the Choi criteria compared to pathological effects,and to observe the consistency between them.Methods:The clinicopathological data of 37 patients,who underwent preoperative treatment with targeted imatinib therapy for GIST,were retrospectively analyzed.Survival ana-lysis of the Choi criteria and pathological effects was conducted using the Kaplan-Meier method and Log-rank test.The consistency between the Choi criteria and the pathological effects was assessed using Spearman's correlation and Kappa tests.Results:The median preoperative treatment duration for the 37 patients was 10 months(range,2-36 months).According to the Choi criteria,there were no cases of complete response(CR),26 cases of partial response(PR),five cases of stable disease(SD),and six progressive disease(PD)cases.The difference in overall survival(OS)between the effective group(CR+PR)and the ineffective group(SD+PD)was statistically significant(P<0.01).Pathologic-al effects were evaluated as one complete effect,11 high effects,18 partial effects,and seven zero effect cases.The OS significantly differed between the effective(full effect+high effect+partial effect)and ineffective(zero effect)groups was statistically significant(P<0.01).The Choi showed moderate consistency with the pathological effects(r=0.592,P<0.01)with a(κappa=0.566).Conclusions:The Choi criteria were moderately correlated and consistent with the pathological effects.Both can be used to evaluate the efficacy and prognosis of preoperative targeted therapy for GIST.The combined use of these two criteria has better clinical application value than that of either alone.

BACKGROUNDNon-alcoholic fatty liver disease (NAFLD) is a complex disorder and has been closely linked to obesity. The fat mass and obesity-associated (FTO) gene is a newly discovered gene related to obesity, which enhances oxidative stress and lipogenesis in NAFLD. The forkhead transcription factor O1 (FoxO1) is another important gene involved in NAFLD, which causes lipid disorders when insulin resistance appears in the liver. However, the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated. This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.

METHODSThis study includes two parts referred to as animal and cell experiments. Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), total triglyceride (TG), total cholesterol (TC), alkaline phosphatase (ALP), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were measured. Immunohistochemical analysis was used to detect the expression and histological localization of FTO, FoxO1, and adenosine monophosphate (AMP)-activated protein kinase (AMPK). The L02 cells were exposed to high fat for 24, 48, or 72 hours. Oil red O staining was used to detect intracellular lipid droplets. Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.

RESULTSAt the end of 10 weeks, ALP, ALT, AST, and LDL were significantly increased (P < 0.01), while TC and TG were also significantly higher (P < 0.05). In addition, HDL was significantly decreased (P < 0.05). The FTO and FoxO1 proteins were weakly expressed in the control group, but both FTO and FoxO1 were expressed significantly higher (P < 0.01) in the experimental group, and the expression of the two factors was significantly correlated. AMPK in the high-fat group showed a low level of correlation with FTO, but not with FoxO1. Oil Red O staining results showed that the cells cultured in 50% fetal bovine serum for 24, 48, or 72 hours exhibited steatosis. FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group (P < 0.01). The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours (P < 0.05).

CONCLUSIONSA high-fat diet leads to higher expression of FTO, phosphorylation of FoxO1, and decreased phosphorylation of AMPK. These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.

Alanine Transaminase ; genetics ; metabolism ; Animals ; Aspartate Aminotransferases ; genetics ; metabolism ; Cholesterol ; metabolism ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; genetics ; metabolism ; Lipoproteins, HDL ; metabolism ; Lipoproteins, LDL ; metabolism ; Liver ; metabolism ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; metabolism ; pathology ; Obesity ; metabolism ; pathology ; Triglycerides ; metabolism

Background Non-alcoholic fatty liver disease (NAFLD) is a complex disorder and has been closely linked to obesity.The fat mass and obesity-associated (FTO) gene is a newly discovered gene related to obesity,which enhances oxidative stress and tipogenesis in NAFLD.The forkhead transcription factor O1 (FoxO1) is another important gene involved in NAFLD,which causes lipid disorders when insulin resistance appears in the liver.However,the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated.This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.Methods This study includes two parts referred to as animal and cell experiments.Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model.Aspartate aminotransferase (AST),alanine aminotransferase (ALT),total triglyceride (TG),total cholesterol (TC),alkaline phosphatase (ALP),high-density lipoprotein (HDL),and low-density lipoprotein (LDL) were measured.Immunohistochemical analysis was used to detect the expression and histological localization of FTO,FoxO1,and adenosine monophosphate (AMP)-activated protein kinase (AMPK).The L02 cells were exposed to high fat for 24,48,or 72 hours.Oil red O staining was used to detect intracellular lipid droplets.Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.Results At the end of 10 weeks,ALP,ALT,AST,and LDL were significantly increased (P ＜0.01),while TC and TG were also significantly higher (P ＜0.05).In addition,HDL was significantly decreased (P ＜0.05).The FTO and FoxO1 proteins were weakly expressed in the control group,but both FTO and FoxO1 were expressed significantly higher (P ＜0.01) in the experimental group,and the expression of the two factors was significantly correlated.AMPK in the high-fat group showed a low level of correlation with FTO,but not with FoxO1.Oil Red O staining results showed that the cells cultured in 50％ fetal bovine serum for 24,48,or 72 hours exhibited steatosis.FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group (P ＜0.01).The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours (P ＜0.05).Conclusions A high-fat diet leads to higher expression of FTO,phosphorylation of FoxO1,and decreased phosphorylation of AMPK.These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.