Background Di(2-ethylhexyl) phthalate (DEHP) is the most prevalent environmental endocrine disruptor among phthalate acid esters (PAEs) worldwide. Previous studies have indicated that exposure to DEHP may disrupt glucose metabolism. Objective To investigate the impact of DEHP on glucose homeostasis in rats, focusing on oxidative stress-induced impairment of autophagy in islet β cells. Methods Forty male SD rats were randomly assigned to four groups, receiving DEHP doses of 0, 187, 375, and 750 mg·kg⁻¹ for 12 weeks. Oral glucose tolerance (OGTT) and insulin tolerance tests (ITT) were conducted 24 h after the final exposure. Pancreatic microstructural alterations were assessed using hematoxylin and eosin (HE) staining and transmission electron microscopy (TEM). Commercial ELISA kits were employed to quantify the levels of insulin, adenosine triphosphate (ATP), and adenosine monophosphate (AMP) in rat serum, as well as the protein expression level of activated caspase-3 in pancreatic tissue. Additionally, commercial microplate kits were utilized to measure the concentration of reduced glutathione (GSH) in serum, the activity of superoxide dismutase (SOD) using water-soluble tetrazolium salt-1, the content of malondialdehyde (MDA) by thiobarbituric acid method, and the level of reactive oxygen species (ROS) in pancreatic tissue by chemical fluorescence method. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure sequestosome1 (SQSTM1/p62), Beclin1, microtubule-associated protein 1 light chain 3 (LC3), and cysteinyl aspartate specific proteinase-8 (Caspase-8) mRNA levels. Western blot analysis was applied to detect the protein relative expression levels of p62, Beclin-1, LC3-I, LC3 II, AMPK, p-AMPK, mTOR, p-mTOR, ULK1, and Caspase-8. Results Compared to the 0 mg·kg⁻¹ DEHP group, the 750 mg·kg⁻¹ DEHP group exhibited a significant increase in fasting blood glucose levels at 2, 4, 6, and 12 weeks (P<0.05). The OGTT showed that, following high-glucose gavage, the 187 mg·kg⁻¹ DEHP group had elevated blood glucose at 30 min (P<0.05), the 375 mg·kg⁻¹ DEHP group showed increased glucose levels at 15, 30, and 180 min (P<0.05), and the 750 mg·kg⁻¹ DEHP group exhibited elevated levels at 15, 30, 60, and 180 min (P<0.05). The 375 and 750 mg·kg⁻¹ DEHP groups demonstrated significantly increased OGTT area under the curve (AUC) values (P<0.05). In contrast, ITT results indicated no significant differences in blood glucose levels or AUC among the DEHP exposure groups at all time points (P>0.05). Compared to the 0 mg·kg⁻¹ DEHP group, the 750 mg·kg⁻¹ DEHP group exhibited significantly higher HOMA-IR levels and markedly lower HOMA-ISI values (P<0.05). HE and TEM showed that in each DEHP exposure group, the number of islet cells decreased, the islet area reduced, and chromatin condensation occurred. The endocrine granules in the cytoplasm of islet β cells decreased, and there were varying degrees of widening of the nuclear membrane gap, flattening and expansion of the Golgi complex, and expansion of the endoplasmic reticulum. Ribosome separation was observed, and autophagosomes were visible. In the 375 and 750 mg·kg⁻¹ DEHP groups, the mitochondria were deformed to varying degrees, and some cristae structures disappeared, presenting vacuolization. Moreover, the chromatin condensation in the nuclei was more severe in the 750 mg·kg⁻¹ DEHP group. The serum SOD activity was significantly elevated in the 750 mg·kg⁻¹ DEHP group (P<0.05). Both the 375 mg·kg⁻¹ and 750 mg·kg⁻¹ DEHP groups exhibited a significant increase in the relative ROS content in pancreatic tissue (P<0.05). In DEHP-treated groups, the MDA content increased (P<0.05), while the GSH content decreased (P<0.05). Additionally, in the 750 mg·kg⁻¹ DEHP group, the AMP/ATP ratio in serum was significantly raised (P<0.05), and the expression of cleaved Caspase-3 protein in pancreatic tissue was also significantly increased (P<0.05). The relative mRNA levels of p62, Beclin-1, LC3, and Caspase-8 in the pancreatic tissue of rats exposed to DEHP were significantly elevated (P<0.05). The relative expression levels of p-AMPK/AMPK, p-ULK1/ULK1, and Beclin-1 proteins in the DEHP-treated groups were significantly increased (P<0.05). In the 375 mg·kg⁻¹ and 750 mg·kg⁻¹ DEHP treatment groups, the relative expression levels of p62, LC3 II/LC1, and Caspase-8 proteins were significantly increased (P<0.05), while the relative expression level of p-mTOR/mTOR was significantly decreased (P<0.05). Conclusion DEHP can disrupt glucose homeostasis by inducing oxidative stress, which subsequently activates autophagy via the ROS/AMPK/ULK1 pathway, impairing autophagic flux and promoting apoptosis of islet β cells, ultimately decreasing their function and number.

Background Di(2-ethylhexyl) phthalate (DEHP) is the highest consumed and the most widely used phthalic acid ester, their effects on lipid metabolism have attracted the attention of many scholars. However, the associated mechanism is still unclear. Objective To observe the effect of DEHP on lipid metabolism in rats, probe its possible mechanism, and provide a research basis for the effect of DEHP on human lipid metabolism. Methods Forty healthy male SD rats were randomly divided into 4 groups: solvent control (0 mg·kg⁻¹ DEHP), low DEHP (187 mg·kg⁻¹), medium DEHP (375 mg·kg⁻¹), and high DEHP (750 mg·kg⁻¹) groups. DEHP was administered by oral gavage for 6 d per week, consecutively 8 weeks. The rats were weighed once a week during the exposure period. At 24 h after the last exposure, the rats were anesthetized with 20% urethane and sacrificed by apical puncture. Rat livers were harvested and weighed before hematoxylin-eosin (HE) histopathological observation. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA levels of lipid metabolism-related genes Janus kinase 3 (JAK3), signal transducer and activator of transcription 5b (STAT5b), and peroxisome proliferator-activated receptor γ (PPARγ) in liver, and Western blot was used to detect the expression levels of lipid metabolism-related proteins JAK3, STAT5b, and PPARγ in liver. Results Compared with the control group, there was no significant difference in the body weight gain of the rats in each group (P>0.05). The liver organ coefficients of the DEHP exposure groups were higher than that of the control group (P<0.001), and increased with higher DEHP dosages. The level of high-density lipoprotein cholesterol (HDL-C) in serum decreased in all DEHP exposure groups (P<0.05), and the level of low-density lipoprotein cholesterol (LDL-C) in serum increased in the high DEHP group (P<0.05). The results of liver histopathological morphology showed that the hepatocytes of each DEHP group were enlarged and edematous in varying degrees, with loose stroma and irregular arrangement of cells, which were manifested as inflammatory cell infiltration and fatty degeneration of liver cells. Compared to the control group, the mRNA levels of JAK3, STAT5b, and PPARγ in liver tissues of rats in each DEHP group decreased (P<0.001). Compared to the control group, the relative expression levels of JAK3 in each DEHP group decreased (P<0.05), and the relative expression levels of STAT5b and PPARγ in the medium and high DEHP groups decreased (P<0.05). Conclusion DEHP exposure can induce abnormal lipid metabolism in rats, and the mechanism may be related to DEHP inhibiting the activation of JAK3/STAT5b/PPARγ signaling pathway.

Objective To analyze the differences in the initial stability of an acetabular cup with a Voronoi polyhedral porous structure and a solid acetabular cup and to explore the impact of the Voronoi polyhedral porous layer on the initial stability of the acetabular cup,as well as its role in preventing loosening and dislocation.Methods Voronoi polyhedral porous scaffold structures with 60％and 70％porosities were designed using the Grasshopper software.Specimens of porous acetabular cups with 60％and 70％porosities and solid acetabular cups were manufactured using selective laser melting technology.Lever tests on the acetabular cups were conducted using polyurethane block models under identical conditions,and the maximum lever-out moment,angular displacement,and interface stiffness of the three groups of specimens were analyzed and compared.Results Under the condition of no significant differences in the compression force,for porous acetabular cups with porosities of 60％and 70％,the maximum lever-out moment increased by 278.82％and 320.56％,the angular displacement increased by 194.04％and 269.23％,respectively,and the interface stiffness increased by 18.58％and 7.88％,respectively,compared with that of solid acetabular cups.After the lever-out tests were completed,significant wear was observed within the polyurethane block hemisphere cavity using the porous acetabular cups.Conclusions The initial stability indicators of acetabular cups with a Voronoi polyhedral porous structure were higher than those of solid acetabular cups,indicating that the Voronoi polyhedral porous layer can enhance the initial stability of the acetabular cup.These results provide a reference for designing and selecting acetabular components.

Background Di(2-ethylhexyl)phthalate (DEHP) and dibutyl phthalate (DBP) are representative environmental endocrine disruptors of phthalate esters (PAEs). Some studies have shown that PAEs exposure may have an impact on lipid metabolism. Objective To investigate the effects of DEHP and/or DBP on lipid metabolism in rats and their possible mechanisms of action. Methods Thirty-six weaned healthy SD male rats, 3 weeks old, weighing 50-70 g, were divided into four groups, i.e., a corn oil control group, a DEHP (750 mg·kg⁻¹) group, a DBP (500 mg·kg⁻¹) group, and a DEHP+DBP (750 mg·kg⁻¹+500 mg·kg⁻¹) group. The rats were exposed to DEHP and/or DBP by oral gavage for 8 weeks, and weighed once a week. The rats were anesthetized 24 h after the last dose, and blood was taken from the apical part of the heart. Serum high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), total cholesterol (TC), and triglyceride (TG) were detected. Liver tissues and perigenital adipose tissues were collected, weighed, and one portion of the tissues was fixed in 10% neutral formalin for pathomorphological observation, and another portion was used for mRNA detection of lipid metabolism-related genes such as Janus kinase 3 (JAK3), signal transducer and activator of transcription 5b (STAT5b), and peroxisome proliferator-activated receptor γ (PPARγ). Results During the DEHP and/or DBP exposure period, the rats in all groups were free to eat and drink without death or injury observed. Compared with the control group: The body weight gain in the DEHP+DBP group was lower at all time points from the 2nd week onwards (P<0.05); the liver organ coefficients of the DEHP and the DEHP+DBP groups were higher (P<0.05); the serum LDL-C levels in the DEHP and the DBP groups were higher (P<0.05). Compared with the DEHP+DBP group: The body weight gains in the DEHP group at the 2nd, 4th, 5th, and 8th weeks were higher (P<0.05), and the body weight gains in the DBP group were higher at all time points except the 1st week (P<0.05); the liver organ coefficients in the DEHP group and the DBP group were lower (P<0.05); the serum TG level in the DEHP group was higher(P<0.05), and the serum LDL-C levels in the DEHP and the DBP groups were higher (P<0.05). The pathomorphological results of liver tissues showed that the hepatocytes in the DEHP, DBP, and DEHP+DBP groups were disordered with loss of cord-like arrangement, swelling (suggesting change of cell proliferation), and presented bilirubin pigmentation. The pathomorphological results of rat perigenital adipose tissues showed had irregular alignment, sizes, and arrangement of adipocyte in the DEHP, DBP, and DEHP+DBP groups. The results of rat liver lipid metabolism-related gene mRNA levels showed that the liver JAK3, STAT5b, and PPARγ mRNA levels in the DEHP, DBP, and DEHP+DBP groups were lower than those in the control group (P<0.05); the rat liver PPARγ mRNA levels in the DEHP and DBP groups were lower than those in the DEHP+DBP group (P<0.05). Conclusion DEHP and/or DBP can inhibit the increase of body weight to varying degrees, induce inflammatory damage to liver tissues, and cause abnormal lipid metabolism in rats, and the associated mechanism may be related to inhibiting the activation of JAK3/STAT5b/PPARγ signaling pathway in rat liver tissues.

OBJE CTIVE To prepare and characterize evodiamine phospholipid complex self-microemulsifying drug delivery system（EVO-PC-SMEDDS），and to investigate its gastric mucosal permeability. METHODS EVO-PC-SMEDDS was prepared ， and particle size ，polydispersity（PDI）and Zeta potential were tested ，and microscopic observation was carried out. The stability of EVO-PC-SMEDDS in simulated gastric liquid with different pH （1.2，2.0，4.0，7.0）was investigated. The entrapment efficiency and drug-loading amount of the preparation were determined ，and the in vitro release was investigated. The gastric mucosal permeability of EVO-PC-SMEDDS was studied by combining rat gastric mucosal tissue and Ussing Chamber technology. RESULTS The particle size of EVO-PC-SMEDDS was （53.63±1.51）nm，PDI and Zeta potential were 0.217±0.017 and （－12.20±0.15）mV，entrapment efficiency was （95.25±0.97）％ and drug-loading amount was （19.30±1.21）mg/g. EVO-PC- SMEDDS exhibited a uniformly dispersed round spherical shape under transmission electron microscope. Stability experiments showed that EVO-PC-SMEDDS exhibited no significant change in particle size ，PDI and Zeta potential under the simulated gastric fluid with different pH and showed excellent stability. Results of in vitro release test showed that compared with evodiamine （EVO），in vitro accumulative release of EVO-PC-SMEDDS were enhanced 6.83-fold，which was in line with the first-order kinetic release model. Results of gastric mucosal permeability showed that gastric mucosal permeation transport ，permeation rate ， permeation flux and area under curve of cumulative permeability of EVO-PC-SMEDDS were higher than those of EVO ， respectively. CONCLUSIONS EVO-PC-SMEDDS is prepared N successfully and shows good stability. It could significantly improve the release behavior and gastric mucosal permeability of EVO.

OBJECTIVE：To optimize the form ulation of Zuojin pectin c apsules，and to prepare modern Zuojin pectin capsules with protective effects against gastric ulcers. METHODS ：The formulation of Zuojin pectin capsules was optimized with orthogonal test with the contents of pectin ，soluble starch and dextrin as factors ，using formability ，moisture absorption and flow ability as indicators. Zuojin pectin capsule was prepared by wet granulation filling method with Zuojin extract powder as raw material. The contents of palmatine hydrochloride ，berberine hydrochloride ，evodiamine and rutaecarpin were evaluated by HPLC. Basket method was used to investigate the release behavior of the capsule in 0.1 mol/L HCl solution. The gastric ulcer model of rats was established by intragastric administration of 75％ ethanol. Gastric ulcer index ，the inhibition rate of gastric ulcer and the pathological sections were used as indexes to investigate the protective effect of Zuojin pectin capsules （the doses were 54，108， 216 mg/kg）on gastric ulcer. RESULTS ：The optimal formulation of Zuojin pectin capsules included 45％ pectin，12％ soluble starch，27％ dextrin and 1％ xylitol. Results of in vitro drug ， release showed that palmatine hydrochloride and berberine， hydrochloride in Zuojin pectin capsules released 53.76％ and No.54.82％ respectively within 1 h，completely released at about 8 h， and conformed to the zero-order release behavior. 2492109374@qq.com Different doses of Zuojin pectin capsule could improve the ulcer injury of gastric tissue in gastric ulcer model rats to different extent ，and significantly reduced the gastric ulcer index（P＜0.01），significantly increased the inhibition rate of gastric ulcer and the percentage of positive expression area of Schiff ’s iodate staining （P＜0.01）. CONCLUSIONS ：Zuojin pectin capsule with protective effect on gastric ulcer and certain sustained- release effect is successfully prepared.

Objective To investigate the effects of Paraquat on human embryonic lung fibroblasts (MRC5) and explore the role of transforming growth factor-β1/connective tissue growth factor signaling pathway in paraquat-induced pulmonary fibrosis.Methods MRC5 cells were cultured with different concentration of PQ (0,12.5,25,50,100,200,400 μ mol/L) for 24 h.The viability of cells was measured by MTT.The protein level of TGF-β1 were analyzed by ELISA after PQ treatment (0,25,50,I00 μmol/L).To examine whether TGF-β1/CTGF signaling pathway was involved in paraquat-induced cytotoxicity,cells was divided into 6 groups:(1) control;(2) 25 μ mol/L PQ group;(3) 50 μ mol/L PQ group;(4) 100 μmol/L PQ group;(5) TGF-β1 positive control group (50 μmol/L rhTGF-β1);(6)stimulate group (100 μmol/L PQ+50 μ mol/L TGF-β1).The protein levels of p-Smad2,p-Smad3 and CTGF were assayed by western blot.The mRNA level of CTGF was assayed by real time RT-PCR.Results MTT showed that cell viability decreased with increasing PQ concentration (P＜0.05).The protein expression of TGF-β1 treated with PQ (25,50,100 μmol/L) significantly increased compared with control in a dose-independent manner(P＜0.05).Exposure to PQ (25,50,100 μmol/L) induced increase of protein levels of p-Smad2 and p-Smad3.Noteworthy,the expression of p-Smad2 and p-Smad3 were dramatically increased following PQ plus TGF-β1 stimulation (P＜0.05).Exposure to PQ (50,100 μmol/L) induced increase of CTGF protein expression and similar greatly increase following PQ plus TGF-β1 stimulation (P＜0.05).Real time RT-PCR showed CTGF mRNA in all groups also significantly up-regulated compared with control (P＜0.05).Conclusion TGF-β1 regulates the expression of target gene CTGF to exhibit its profibrogenic effects by activating TGF-β1/Smad signaling pathway in PQ-induced pulmonary fibrosis.

Objective To investigate the association between occupational stress and activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in patients with metabolic syndrome.Methods A case-control study was performed.According to inclusion and exclusion criteria,among the staff members of enterprises and public institutions aged 20～60 years who underwent physical examination in The Affiliated Hospital of Ningxia Medical University and The People's Hospital of Wuzhong from October 2011 to October 2012,622 patients with metabolic syndrome who did not have a blood relationship with each other were enrolled as case group,and 600 healthy staff members who also did not have a blood relationshipwith each otherwere enrolled as control group.Questionnaire investigation,chronic occupational stress investigation,physical examination,and laboratory tests were performed for all subjects.Results Compared with the control group,the case group had significantly higher serum levels and abnormal rates of AST and ALT (t=-4.338 and-5.485,x2=11.168 and 34.302,all P＜0.05).There were no significantdifferences in the serum level and abnormal rate of AST between the subgroups with different occupational stresses in both groups (F=2.192 and 2.567,x2=2.694 and 5.402,all P＞0.05),but there were significant differencesbetween the subgroups in all subjects (F=5.005,x2=6.398,all P＜0.05).There were no significant differences in the serum level and abnormal rate of ALT between thesubgroups with different occupational stresses in the case group,the control group,and all subjects (F=0.845,0.450,and 1.416,x2=2.564,1.344,and 3.147,all P＞0.05).The partial correlation analysis showed that the total score of occupational stress was positively correlated withthe serum level of AST (r=0.071,P＜0.05) and was not correlatcd with the serum level of ALT(r=-0.044,P＞0.05),and that the serum level of AST was positively correlated with that of ALT (r=0.736,P＜0.05).After the adjustment for age,sex,nationality,smoking,drinking,marital status,and degree of education,the total score of occupational stress was positively correlated with the serum level of AST (r=0.069,P＜0.05) and was not correlated with the serum level of ALT (r=-0.042,P＞0.05),and the serum level of AST was positively correlated with that of ALT(r=0.730,P＜0.05).The multiple linear regression analysis showed that the serum level of AST increased with the increasing occupational stress (b=0.131,P=0.013).Conclusion Occupational stress is associated with increased serum level of AST,and the serum level of AST increases with the increasing occupational stress.Increased risk of metabolic syndrome caused by occupational stress may be associated with the increased activity of AST caused by occupational stress.

Objective To investigate the effects of Paraquat on human embryonic lung fibroblasts (MRC5) and explore the role of transforming growth factor-β1/connective tissue growth factor signaling pathway in paraquat-induced pulmonary fibrosis.Methods MRC5 cells were cultured with different concentration of PQ (0,12.5,25,50,100,200,400 μ mol/L) for 24 h.The viability of cells was measured by MTT.The protein level of TGF-β1 were analyzed by ELISA after PQ treatment (0,25,50,I00 μmol/L).To examine whether TGF-β1/CTGF signaling pathway was involved in paraquat-induced cytotoxicity,cells was divided into 6 groups:(1) control;(2) 25 μ mol/L PQ group;(3) 50 μ mol/L PQ group;(4) 100 μmol/L PQ group;(5) TGF-β1 positive control group (50 μmol/L rhTGF-β1);(6)stimulate group (100 μmol/L PQ+50 μ mol/L TGF-β1).The protein levels of p-Smad2,p-Smad3 and CTGF were assayed by western blot.The mRNA level of CTGF was assayed by real time RT-PCR.Results MTT showed that cell viability decreased with increasing PQ concentration (P＜0.05).The protein expression of TGF-β1 treated with PQ (25,50,100 μmol/L) significantly increased compared with control in a dose-independent manner(P＜0.05).Exposure to PQ (25,50,100 μmol/L) induced increase of protein levels of p-Smad2 and p-Smad3.Noteworthy,the expression of p-Smad2 and p-Smad3 were dramatically increased following PQ plus TGF-β1 stimulation (P＜0.05).Exposure to PQ (50,100 μmol/L) induced increase of CTGF protein expression and similar greatly increase following PQ plus TGF-β1 stimulation (P＜0.05).Real time RT-PCR showed CTGF mRNA in all groups also significantly up-regulated compared with control (P＜0.05).Conclusion TGF-β1 regulates the expression of target gene CTGF to exhibit its profibrogenic effects by activating TGF-β1/Smad signaling pathway in PQ-induced pulmonary fibrosis.

Objective To investigate the association between occupational stress and activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in patients with metabolic syndrome.Methods A case-control study was performed.According to inclusion and exclusion criteria,among the staff members of enterprises and public institutions aged 20～60 years who underwent physical examination in The Affiliated Hospital of Ningxia Medical University and The People's Hospital of Wuzhong from October 2011 to October 2012,622 patients with metabolic syndrome who did not have a blood relationship with each other were enrolled as case group,and 600 healthy staff members who also did not have a blood relationshipwith each otherwere enrolled as control group.Questionnaire investigation,chronic occupational stress investigation,physical examination,and laboratory tests were performed for all subjects.Results Compared with the control group,the case group had significantly higher serum levels and abnormal rates of AST and ALT (t=-4.338 and-5.485,x2=11.168 and 34.302,all P＜0.05).There were no significantdifferences in the serum level and abnormal rate of AST between the subgroups with different occupational stresses in both groups (F=2.192 and 2.567,x2=2.694 and 5.402,all P＞0.05),but there were significant differencesbetween the subgroups in all subjects (F=5.005,x2=6.398,all P＜0.05).There were no significant differences in the serum level and abnormal rate of ALT between thesubgroups with different occupational stresses in the case group,the control group,and all subjects (F=0.845,0.450,and 1.416,x2=2.564,1.344,and 3.147,all P＞0.05).The partial correlation analysis showed that the total score of occupational stress was positively correlated withthe serum level of AST (r=0.071,P＜0.05) and was not correlatcd with the serum level of ALT(r=-0.044,P＞0.05),and that the serum level of AST was positively correlated with that of ALT (r=0.736,P＜0.05).After the adjustment for age,sex,nationality,smoking,drinking,marital status,and degree of education,the total score of occupational stress was positively correlated with the serum level of AST (r=0.069,P＜0.05) and was not correlated with the serum level of ALT (r=-0.042,P＞0.05),and the serum level of AST was positively correlated with that of ALT(r=0.730,P＜0.05).The multiple linear regression analysis showed that the serum level of AST increased with the increasing occupational stress (b=0.131,P=0.013).Conclusion Occupational stress is associated with increased serum level of AST,and the serum level of AST increases with the increasing occupational stress.Increased risk of metabolic syndrome caused by occupational stress may be associated with the increased activity of AST caused by occupational stress.