Objective : To investigate the protective effect of dimethyl fumarate(DMF) on maternal intrahepatic cholestasis(ICP) during pregnancy induced by di(2-ethylhexyl) phthalate(DEHP) exposure and its mechanism. Methods : Thirty-two 8-week-old female institute of cancer research(ICR) mice were randomly divided into 4 groups: Ctrl group, DEHP group, DMF group and DEHP+DMF group. DEHP and DEHP+DMF groups were treated with DEHP(200 mg/kg) by gavage every morning at 9:00 a.m. DMF and DEHP+DMF groups were treated with DMF(150 mg/kg) from day 13 to day 16 of gestation by gavage. After completion of gavage on day 16 of pregnancy, maternal blood, maternal liver, placenta, and amniotic fluid were collected from pregnant mice after a six-hour abrosia. The body weight of the mother rats and the body weight of the fetus rats were sorted and analyzed; the levels of total bile acid(TBA), alkaline phosphatase(ALP), aspartate aminotransferase/alanine aminotransferase(AST/ALT) in serum and TBA in liver, amniotic fluid and placenta were detected by biochemical analyzer; HE staining was used to observe the pathological changes of liver tissue; Quantitative reverse transcription PCR(RT-qPCR) was used to detect the expression levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-1, IL-18 and NOD-like receptor thermal protein domain associated protein 3(NLRP3) in the liver; Western blot was used to detect the expression of the nuclear factor KappaB(NF-κB) and NLRP3. Results : Compared with the control group, the body weight of the DEHP-treated dams and pups decreased(P<0.05); the levels of TBA, ALP, AST/ALT in the serum of dams and the levels of TBA in the liver, amniotic fluid, and placenta of dams increased(P<0.05); the histopathological results showed that liver tissue was damaged, bile ducts were deformed, and there was inflammatory cell infiltration around them; the levels of inflammation-related factors TNF-α, IL-6, IL-1, IL-18 and NLRP3 transcription in maternal liver increased(P<0.05); the expression of NF-κB and NLRP3 protein in maternal liver significantly increased( P<0. 05). Compared with the DEHP group,the body weight of both dams and fetuses significantly increased in DEHP + DMF group( P<0. 05); the levels of TBA,ALP,AST/ALT in the serum of dams and amniotic fluid of fetuses decreased( P<0. 05); the degree of liver lesions was improved; the transcription levels of inflammation-related factors TNF-α,IL-6,IL-1,IL-18 and NLRP3 in maternal liver decreased( P<0. 05); the expression of NF-κB and NLRP3 protein in maternal liver significantly decreased( P<0. 05). Conclusion DMF can effectively protect the DEHP exposure to lead to female ICP,and its mechanism may be through inhibiting the NF-κB/NLRP3 pathway and reducing liver inflammation.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Objective : To establish a method for measuring major organic acids in the tricarboxylic acid cycle using a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) system, and to investigate the changes in six tricarboxylic acid cycle organic acids(fumaric acid, malic acid, succinic acid, α-ketoglutaric acid, cis-aconitic acid, and citric acid) in the serum, liver, and placenta of mice exposed to di(2-ethylhexyl) phthalate(DEHP) during pregnancy. Methods : The serum, liver and placental samples from pregnant mice were processed and eluted through a Waters ACQUITY UPLC BEH Amide Column(130 Å, 1.7 μm, 2.1 mm × 150 mm) using a gradient elution program. Mobile phase A comprised an aqueous solution of 10 mmol/L ammonium acetate and 5 μmol/L methanephosphonic acid, while mobile phase B consisted of a 90% acetonitrile aqueous solution containing 10 mmol/L ammonium acetate and 5 μmol/L methanephosphonic acid, with a flow rate maintained at 0.35 ml/min. The mass spectrometry detection system utilized an electrospray ionization technique with negative ion mode for multiple reaction monitoring. Results : The correlation coefficients of the standard curves for the six tricarboxylic acid cycle organic acid metabolites were all above 0.996 within the quantitative range. The method's accuracy ranged from 97.14% to 108.26%, with inter-day and intra-day precision relative standard deviation between 1.35% and 6.73%. The matrix effect was between 93.29% and 107.47%, and the extraction recovery rate ranged from 94.82% to 112.57%. Analysis of six tricarboxylic acid cycle organic acids in the liver, serum, and placenta of DEHP-exposed mice during pregnancy showed significant reductions in fumaric acid, malic acid, α-ketoglutaric acid, cis-aconitic acid, and citric acid compared to the control group(P<0.05). Conclusion The HPLC-MS/MS method established in this study for detecting six tricarboxylic acid cycle organic acids in the serum, liver, and placenta of DEHP-exposed pregnant mice is stable, highly sensitive and selective. Prenatal DEHP exposure induced alterations in the levels of tricarboxylic acid(TCA) cycle organic acid metabolites in the liver, serum, and placenta of mice, suggesting that DEHP exposure during pregnancy may interfere with mitochondrial TCA cycle processes. These findings indicate potential value in the diagnosis and treatment of diseases associated with prenatal DEHP exposure.

Objective:To explore the feasibility of preparing an iodine-coated titanium plate with potassium iodide and verify its antibacterial performance.Methods:Iodine was coated onto the surface of a titanium plate in electrolyte of potassium iodide using the electrophoretic deposition method. The signs and composition of the surface of the iodine-coated titanium plate were observed by scanning electron microscopy and energy dispersive spectroscopy. The experiment was conducted in a control group and 3 antibacterial test groups. The control group consisted of 10 titanium plates which had been pretreated but not loaded with iodine; the 3 experimental groups also had in each 10 titanium plates which had been pretreated and loaded with iodine in the electrolytes of concentrations of 1,000 mg/L, 2,000 mg/L and 4,000 mg/L, respectively. The antimicrobial tests in vitro were conducted with standard strains of staphylococcus aureus [1×10 6 Colony-Forming Units (CFU)/mL ATCC25923] to determine the antibacterial property of the plates. Results:The iodine-coated titanium plates appeared grey and their surface was evenly covered with a flat coating with no collapse. The scanning electron microscopy observed on the surface of the iodine-coated titanium plates an iodine coating with scattered irregular collapses in different sizes. The iodine content was 0 mass%, 5.10 mass%, 10.32 mass% and 15.05 mass%, respectively, in the control, 1,000 mg/L, 2,000 mg/L and 4 000 mg/L groups under the energy dispersive spectroscopy. Their counts of in vitro antibacterial colony were 56.00±5.09, 21.40±2.76, 9.10±2.51, and 2.00±1.88, respectively, showing significant differences between groups ( P< 0.05). Conclusions:A titanium plate with a steady and even iodine coating can be prepared by virtue of the electrophoretic deposition method in electrolyte of potassium iodide. The antibacterial property of an iodine-coated titanium plate is superior to that of a titanium plate without iodine coating.

Objective:To explore the influence of organizational management mode and regional medical resource allocation on thrombolysis rate of stroke.Methods:A cross-sectional study was carried out in 61 thrombolytic units distributed at 16 administrative districts of Shanghai to collect information including the number of imaging equipment, neurologists and nurses, hospital organization and management mode, thrombolytic rate, etc. Using SPSS 19.0 statistical software, simple linear regression analysis and chi square test were used to analyze the correlation between related indexes and thrombolysis rate.Results:There was no linear correlation between imaging equipment, human resources and regional thrombolysis rate. The key factor to improve thrombolysis rate was the organizational management of stroke( OR=1.488, 95% CI=1.357-1.631, P<0.001). Conclusions:An effective hospital organization and management model, including the establishment of multi department cooperation, stroke emergency team, stroke green channel, can significantly improve the thrombolysis rate of stroke.

Objective Iodine loading would be carried out on orthopedic titanium Kirschner-wire by electrophoretic deposition.And by this,a kind of orthopedic iodine-coated implant with antibacterial properties was supposed to be developed,that would provide a theoretical basis for the development of clinical orthopedic implants with antibacterial properties.Methods Iodine loading on the surface of titanium Kirschner-wire was carried out by electrophoretic deposition.PVP-I solution as 1 000 ppm,2 000 ppm and 4 000 ppm was prepared respectively (1 ppm=1 mg/kg=1 mg/L).Three kinds of iodine-coated orthopedic titanium Kirschner-wires with different iodine content were prepared by electrophoretic deposition at 200 V voltage for 30 min and then stoved after being washed with distilled water.The surface signs and composition structure of iodine coated titanium Kirschnerwire were studied through scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS).Antibacterial experiments of Iodine coating titanium Kirschner needle were performed in vitro.Antimicrobial test:10 pieces of uncoated titanium Kirschner-wires;10 pieces of iodine-coated titanium Kirschner-wires with 1 000 ppm;10 pieces of iodine-coated titanium Kirschner-wires with 2 000 ppm;10 pieces of iodine-coated titanium wires with 4 000 ppm.Because iodine is easy to sublimate and does not withstand high temperature,all the titanium Kirschner needles are fumigated and disinfected by ethylene oxide (supported by the supply room of our hospital).Titanium wires were immersed respectively in 106 CFUJ/ml (ATCC25923) bacterial suspension of standard Staphylococcus aureus so that bacteria would be fully inoculated on the surface of the titanium wire.The sterilized titanium Kirschner needle was placed in a sterile culture cup and 30 ml suspension was added into the culture cup.The sterilized titanium Kirschner needle was completely immersed in the bacterial solutionand was kept at 37°C for 6 h before taking out.Take out the titanium Kirschner needle slowly,rinse the titanium Kirschner needle with 5 ml sterile PBS buffer,after rinsing the surface of titanium Kirschner needle,transfer the titanium Kirschner needle to the new sterile culture cup and add 5 ml sterile saline until the titanium Kirschner needle is completely immersed.Then put the titanium Kirschner needle into the ultrasonic oscillator to wash the surface of the bacteria fully.After 10 000 times dilution,100 μl was smeared on agar medium for culture (37°C) and the number of colonies was observed and counted 24 h later.Results The surface of three kinds of iodine coating titanium Kirschner-wires was covered with uniform iodized compound coating and had a brown look.The film structure was stable and there was no falling off after ultrasonic cleaning.SEM exhibited that the surface of Titanium Kirschner-wire is covered with a compact coating;the appearance structure is relatively flat and a slight collapse can be seen scattered in the surface of the wire.The results of EDS showed that the iodine content in the surface coating of 3 groups was 4.38 wt％,9.05 wt％ and 14.48 wt％.The bacteria growing on the surface of titanium needle were shaken down by ultrasonic vibration.The CFU counting was carried out after the bacterial solution being diluted,coated and cultured for 24 h.It can be seen that the antibacterial property increases continuously with the increase of iodine coating content.The antibacterial properties of iodine-coated implant which content 14.48 wt％ iodine was more than 1 orders of magnitude than the control group.The results showed that the plants coated with iodine had obvious antibacterial effect.Conclusion The iodine coated titanium wire was prepared by electrophoretic deposition,after being characterized by SEM,EDS and quantitative measurement,the successful loading of iodine was proved,the antibacterial experiment proved that the iodinecoated orthopedic titanium Kirschner-wire has a stronger antibacterial properties than the ordinary titanium Kirschner-wire under laboratory conditions.

Objective: To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion. Methods: Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF. Results: CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05). Conclusions Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.

Objective To observe the efficacy of inhaled tiotropium bromide combined with budesonide/formoterol on reducing the frequency of acute episodes of symptom exacerbation and improving lung function,health status in chronic obstructive pulmonary disease (COPD). Methods Eighty-six patients with COPD were divided into 3 groups, combination group[29 cases, inhaled budesonide/formoterol (160 μg/4.5 μg, twice one day ) and tiotropium bromide ( 18 μg, once one day)], budesonide/formoterol group( 29 cases, 160 μg/4.5 μg, twice one day) and tiotropium bromide group(28 cases, 18 μg, once one day). The treatment continued for 3 months. Results Lung function, symptoms and health status improved obviously in three groups. The forced expiratory volume in one second (FEV1) of combination group after treatment was (1.24±0.18) L , which was improved by 11.7% compared with before treatment. It was significantly higher than that in budesenide/formoterol group and fiotropium bromide group (P < 0.01 ). The rescue medication consumptions and the times of acute episode of combination group were significantly decreased compared with those in the other groups,and there was significant difference (P <0.01). The SGRQ score of combination group was (35.6±13.9) points which was significantly lower than that of budesonide/formoterol group and tiotropium bromide group,and there was significant difference (P < 0.01 ).There was no statistical difference in the adverse events occurred in three groups (P > 0.05 ). Conclusions Combination treatment produces better control of symptoms and lung function and has no greater risk of sideeffects, compared with the treatment of budesonide/formoterol alone and tiotropium bromide alone. The combination treatment should be considered for patients with COPD.