OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.

Objective:To evaluate the effect of gastrogin on AMP-activated protein kinase (AMPK)/transient receptor potential anchor protein 1 (TRPA1) signaling pathway in rats with neuropathic pain.Methods:Thirty-six SPF-grade healthy male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-230 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation+ normal saline group (SHAM group), neuropathic pain+ normal saline group (NP group), and neuropathic pain+ gastrogin group (GAS group). Neuropathic pain was induced by chronic constrictive injury to sciatic nerve under 2% isoflurane anaesthesia. The sciatic nerve was only exposed but not ligated in SHAM group. Gastrogin 100 mg/kg was intraperitoneally injected for 14 consecutive days after developing the model in GAS group, while the equal volume of normal saline was given instead in SHAM and NP groups. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before developing the model (T 0) and 1, 3, 5, 7, 10 and 14 days after developing the model (T 1-6). The rats were anesthetized and sacrificed following the measurement of pain thresholds at T 4 and T 6. The lumbar segment (L 4-6) of the spinal cord was removed for determination of TRPA1 mRNA expression (by quantitative real-time polymerase chain reaction), expression of TRPA1, AMPK and p-AMPK (by Western blot), expression of TRPA1 (by immunofluorescence staining) and expression of tumor necrosis-alpha(TNF-α), interleukin-1beta(IL-1β) and c-fos (by immunohistochemistry). Results:Compared with SHAM group, MWT and TWL were significantly decreased at T 1-6, the expression of TRPA1 mRNA, TRPA1, TNF-α, IL-1β and c-fos was up-regulated, the expression of p-AMPK was down-regulated ( P<0.05), and no significant change was found in AMPK expression in NP group ( P>0.05). Compared with NP group, MWT at T 3-6 and TWL at T 2-6 were significantly increased, the expression of TRPA1 mRNA, TRPA1, TNF-α, IL-1β and c-fos was down-regulated, and p-AMPK expression was up-regulated ( P<0.05), and no significant change was found in AMPK expression in GAS group ( P>0.05). Conclusions:The mechanism by which gastrogin reduces neuropathic pain may be related to modulating the expression of the AMPK/TRPA1 signaling pathway in rats.

Objective Mitochondrial reactive oxygen species(mtROS)could cause damage to pancreatic β-cells,rendering them susceptible to oxidative damage.Hence,investigating the potential of the mitochondria-targeted antioxidant(Mito-TEMPO)to protect pancreatic β-cells from ferroptosis by mitigating lipid peroxidation becomes crucial. Methods MIN6 cells were cultured in vitro with 100 μmol/L sodium palmitate(SP)to simulate diabetes.FerroOrange was utilized for the detection of Fe2+fluorescence staining,BODIPY581/591C11 for lipid reactive oxygen species,and MitoSox-Red for mtROS.Alterations in mitophagy levels were assessed through the co-localization of lysosomal and mitochondrial fluorescence.Western blotting was employed to quantify protein levels of Acsl4,GPX4,FSP1,FE,PINK1,Parkin,TOMM20,P62,and LC3.Subsequently,interventions were implemented using Mito-TEMPO and Carbonyl cyanide 3-chlorophenylhydrazone(CCCP)to observe changes in ferroptosis and mitophagy within MIN6 cells. Results We found that SP induced a dose-dependent increase in Fe2+and lipid ROS in MIN6 cells while decreasing the expression levels of GPX4 and FSP1 proteins.Through bioinformatics analysis,it has been uncovered that mitophagy assumes a crucial role within the ferroptosis pathway associated with diabetes.Additionally,SP decreased the expression of mitophagy-related proteins PINK1 and Parkin,leading to mtROS overproduction.Conversely,Mito-TEMPO effectively eliminated mtROS while activating the mitophagy pathways involving PINK1 and Parkin,thereby reducing the occurrence of ferroptosis in MIN6 cells.CCCP also demonstrated efficacy in reducing ferroptosis in MIN6 cells. Conclusion In summary,Mito-TEMPO proved effective in attenuating mtROS production and initiating mitophagy pathways mediated by PINK1 and Parkin in MIN6 cells.Consequently,this decreased iron overload and lipid peroxidation,ultimately safeguarding the cells from ferroptosis.

Objective:To investigate the effect of decarbromodiphenyl ether(BDE-209)exposure on the migration ability of triple-negative breast cancer(TNBC)cells and to explore the underlying mechanism.Methods:Human TNBC MDA-MB-231 cells were divided into blank control group and BDE-209 exposure groups(treated with 0.02,0.20,2.00,20.00 and 200.00 ng/mL BDE-209 in high glucose DMEM).Extracellular vehicles(EVs)secreted by MDA-MB-231 cells were isolated by differential ultracentrifugation.Transmission electron microscopy(SEM),nanoparticle tracking analysis(NTA)and Western blotting were performed to characterize the EVs.The effect of the EVs induced by BDE-209 exposure(EVs-BDE-209)on the migration and invasion of MDA-MB-231 cells was detected by wound-healing assay and Transwell test.qRT-PCR was used to measure the miR-221 level in EVs-BDE-209.The expression of MMP9 in MDA-MB-231 cells was determined by Western blotting.Results:Compared with the blank control,BDE-209 exposure increased the tumor cell-derived EVs in dose-dependent manner.The MDA-MB-231 cells co-cultured with EVs released by 200.00 ng/mL BDE-209 exposure showed an 86%increase in cell migration rate,a 1.32-fold higher number of membrane-penetrating cells,a 2.71-fold higher expression level of miR-221,and a 1.62-fold higher expression level of MMP9 compared with the blank control group(all P<0.05).While transfection with anti-miR-221 antibody to decrease miR-221 level in EVs significantly reversed the increased invasion ability of the MDA-MB-231 cells treated with EVs-BDE-209.Conclusion:BDE-209 exposure may promote metastasis potential of MDA-MB-231 cells via EVs-BDE-209 transmitted miR-221.

Objective To investigate the effect of trehalose(Tre)H9C2 cardiomyocytes under oxy-gen-glucose deprivation/reoxygenation(OGD/R)injury and its mechanism of action with the cel-lular model simulating the process of myocardial ischemia/reperfusion injury.Methods H9C2 cells were divided control group,OGD/R group,Tre group(OGD/R+Tre),and combination group(OGD/R+Tre+ML385).MTT assay was used to observe cell proliferation,and lactate de-hydrogenase(LDH)release and Hoechst/propidium iodide staining were employed to evaluate cell membrane damage.Western blot analysis was utilized to detect the expression of nuclear fac-tor erythroid 2-related factor 2(Nrf2)and its downstream related proteins.The generation of re-active oxygen species(ROS)and mitochondrial membrane potential(MMP)were measured to quantify the level of oxidative stress.The expression of apoptosis-related proteins was determined by Western blot analysis.Results In comparison to the control group,the OGD/R group exhibi-ted a significantly reduced cell viability.When compared with the OGD/R group,the intervention of varying concentrations of Tre obviously improved cell viability in a dose-dependent manner(P＜0.01),increased MMP,and up-regulated the expression of glutathione(GSH),Nrf2,hemeo-xygenase 1,and nicotinamide adenine dinucleotide phosphate quinone oxidoreductase 1(NQO1),Bcl-2,and cysteinyl aspartate specific proteinase 3(Caspase-3)(P＜0.05,P＜0.01),and decreased the production of ROS and MDA and the expression of responding element binding protein 1,Bax,Bax/Bcl-2,and cleaved Caspase-3(P＜0.05,P＜0.01).What's more,the combined group ex-hibited significantly higher production of ROS,MDA,increased mRNA levels of TNF-α,IL-1β,and IL-6,and reduced MMP and GSH levels(P＜0.05,P＜0.01),as well as,enhanced expression of Bax,Bax/Bcl-2,and cleaved Caspase-3(1.77±0.08 vs 1.20±0.20,3.41±1.45 vs 0.99±0.15,4.10±1.05 vs 1.79±0.52,P＜0.01),and decreased expression of Bcl-2 and Caspase-3(0.58±0.21 vs 1.23±0.25,0.87±0.25 vs 1.45±0.31,P＜0.01)in comparison with the Tre group.Conclusion Tre can be regarded as an Nrf2 activator that inhibits oxidative stress and apoptosis by activating Nrf2,and thereby ameliorates OGD/R-induced cardiomyocyte injury.

Objective:To evaluate the effect of trehalose on oxygen-glucose deprivation and restoration (OGD/R) injury in H9C2 cells and the role of solute carrier family 7 member 11- (SLC7A11)-glutathione peroxidase 4 (GPX4) signaling pathway.Methods:Well-grown H9C2 cells were divided into 4 groups ( n=24 each) by the random number table method: control group (group C), OGD/R group (group O), OGD/R+ trehalose group (group OT) and OGD/R+ trehalose+ erastin group (group OTE). The cells were normally cultured in group C. In O, OT and OTE groups, the DMEM medium was replaced with EBSS medium, the cells were exposed to 5% CO 2-95% N 2 in an incubator at 37 ℃ for 6 h, and then the medium was replaced with DMEM medium supplemented with 6% fetal bovine serum to restore oxygen and glucose supply for 24 h. In group OT, trehalose at a final concentration of 50 mmol/L was added during restoration of oxygen and glucose supply. In group OTE, the SLC7A11 inhibitor erastin at a final concentration of 10 μmol/L was added at 8 h before oxygen-glucose deprivation, and trehalose at a final concentration of 50 mmol/L was added during restoration of oxygen and glucose supply. The cell viability, lactic dehydrogenase (LDH) activity, contents of glutathione (GSH), malondialdehyde (MDA) and iron, and reactive oxygen species (ROS) levels were measured at 24 h of restoration of oxygen and glucose supply. The expression of SLC7A11, GPX4, long-chain fatty acyl coenzyme A synthetase 4 (ACSL4), and ferritin heavy chain 1 (FTH1) was detected by Western blot. The structure of the mitochondrial morphology was observed with a transmission electron microscope. Results:Compared with group C, the cell viability and GSH content were significantly decreased, the LDH activity, contents of MDA and iron, and ROS level were increased, the expression of SLC7A11, GPX4 and FTH1 was down-regulated, and the expression of ACSL4 was up-regulated in group O ( P<0.05). Compared with group O, the cell viability and GSH content were significantly increased, the LDH activity, contents of MDA and iron, and ROS level were decreased, the expression of SLC7A11, GPX4 and FTH1 was up-regulated, and the expression of ACSL4 was down-regulated in group OT ( P<0.05). Compared with group OT, the cell viability and GSH content were significantly decreased, the LDH activity, contents of MDA and iron, and ROS levels were increased, the expression of SLC7A11, GPX4 and FTH1 was down-regulated, and the expression of ACSL4 was up-regulated in group OTE ( P<0.05). Conclusions:Trehalose can inhibit ferroptosis by activating the SLC7A11-GPX4 signaling pathway, thereby attenuating OGD/R injury in H9C2 cells.

Diet management is the key part in the prevention and treatment of chronic kidney disease (CKD). A plant-based diet is a diet consisting mostly or entirely of foods derived from plants, with little or no animal foods. In recent years, research on plant-based diets for chronic kidney disease has been increasing. Large-scale epidemiological studies and interventional studies consistently suggest that plant-based diets could reduce the risk of CKD and related complications and slow down CKD progression. This article reviews the research progress of plant-based diets in the prevention and control of CKD.

Overweight and obesity are main risk factors for chronic metabolic diseases, and are strongly associated with the increased risk of premature death. Low carbohydrate diet (LCD) has been proven to effectively control body weight and fat mass in overweight and obese patients by short-term (≤6 months) dietary intervention studies. The mechanisms include regulation of energy metabolism, anti-inflammatory, antioxidant, alteration in expression of lipid metabolic-related genes and modulation of intestinal flora. However, the conclusions are inconsistent on whether LCD can cause durable weight loss and reduce the risk of overweight and obesity. This review summarizes the current research progress on the mechanisms, epidemiological studies, intervention studies and potential risks of LCD in controlling overweight and obesity, providing a reference for the future research and clinical application.

Objective To investigate the effects of Bucinnazine Hydrochloride on the pain behavior and the expression of caveolin-1 (Cav-1) in the anterior cingulate cortex of neuropathic pain mice.Methods 64 adult male Kunming mice (20-25g) were divided randomly into 4 groups with 16 in each group:Sham+BH(Bucinnazine Hydrochloride) group,Sham+NS (Normal Saline) group,CCI+ BH group and CCI+ NS group.The corresponding drugs were administered by intraperitoneal injectionfrom the forth day after CCI once a day for three days.Paw thermal withdrawal latency was measured by Hargreaves methods.Mechanicalwithdrawal threshold was assayed by electronic dolorimeter.c-Fos protein in anterior cingulate cortex was detected by immunohistochemistry staining and the expression of t-Cav-1,p-Cav-1was detected by Western blot.Results Bucinnazine Hydrochloride administered by intraperitoneal injection(0.1 mg/10 g,mice) alleviated thermal hyperalgesia and mechanical allodynia of CCI mice.Compared with the forth day (4.92±0.41) s of CCI+BH group,paw withdrawal latency on the fifth day(5.92±0.61) s was increased(P＜0.05),and on the sixth day(7.93±0.91) s and seventh day (9.12±0.69)s were increased more(P＜0.01,P＜0.01).The paw withdrawal mechanical threshold on the sixth and seventh day of CCI+BH group mice((2.54 ±0.41)g,(3.68±0.61)g) were increased significantly (P＜0.01,P＜0.01)compared with the forth day(1.55± 0.31)g.Immunohistochenistry results showed that the expression of c-Fos decreased after treated with Bucinnazine Hydrochloride in the anterior cingulate cortex of CCI mice(P＜0.001).Western Blotting showed that the expression of t-Cav-1 (1.97±0.31) and p-Cav-1 (0.11 ±0.09) in the anterior cingulate cortex of CCI +BH group mice decreased compared with that of in CCI+NS group mice(t-Cav-1:2.87±0.15,p-Cav-1:0.48± 0.09) (P＜0.01,P＜0.01).Conclusion Bucinnazine Hydrochloride can alleviate both thermal hyperalgesia and mechanical allodynia of neuropathic pain of mice,and reduce the expression of c-Fos,t-Cav-1,p-Cav-1 in the anterior cingulate cortex of neuropathic pain mice.