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 investigate the inhibitory effects of magnesium citrate(MgCit)on oxidative stress in chronic renal failure(CRF).Methods SD rats were divided into CRF model group,MgCit groups(375 and 750 mg/kg),normal control group,and MgCit control group(750 mg/kg).The morphology of mitochondria in thoracic artery vascular smooth muscle cells(VSMCs)was observed by transmission electron microscopy.The content of superoxide dismutase(SOD)and malonaldehyde(MDA)in rat aorta and plasma was detected by the kit.The VSMCs were divided into normal control group,CRF model group,and MgCit groups(1.5 and 3 mmol/L).The levels of superoxide anion(DHE)and apoptosis were quantitatively detected by flow cytometry.Results Compared with the control groups,the mitochondria were swollen and the cristae fractured or disappeared in the model group;MgCit intervention could reduce mitochondrial swelling,but not cristae fracture.In the model group,SOD level in aorta and plasma decreased(P＜0.05)while MDA level increased(P＜0.05).MgCit intervention could increase SOD in aorta and plasma,but decrease MDA level(P＜0.05).In the CRF environment,the DHE content of VSMCs and apoptosis in CRF model group increased(P＜0.05).MgCit intervention could decrease DHE content and inhibit apoptosis(P＜0.05).Conclusion MgCit inhibits oxidative stress levels in vivo and in vitro in CRF.

Vaccination for children and adolescents is an effective strategy for preventing and controlling infectious diseases. However， clinical reports showed that vaccination may be associated with epilepsy or seizures， potentially increasing public hesitancy about vaccination， and affecting vaccination coverage. At present， no evidence showed that vaccination could damage the nervous system and cause epilepsy or seizures. In this review， we explore the association of multiple vaccinations in children and adolescents with epilepsy or seizures， and further understand the safety of vaccination， thereby improving vaccine coverage in China.

AIM: To investigate the short-term efficacy of intense pulsed light(IPL)for the comprehensive treatment of blepharokeratoconjunctivitis(BKC).METHODS: A total of 66 patients(66 eyes)diagnosed with BKC in Cangzhou Aier Eye Hospital from December 2020 to December 2021 were selected(All selected the more severe eye for research). They were divided into two groups according to the different clinical treatment, with 30 cases(30 eyes)in control group and 36 cases(36 eyes)in treatment group. Both groups were administrated for 1wk(0.1% fluorometholone eye drops, 0.3% sodium hyaluronate eye drops and tobramycin dexamethasone eye ointment). Ocular surface state was taken as the baseline level. The control group continued to receive drugs(0.1% fluorometholone eye drops and 0.3% sodium hyaluronate eye drops), while the treatment group is required to undergo regular IPL in the hospital on the basis of drug treatment(atomization fumigation+IPL+meibomian gland massage + meibomian margin cleaning+cold compress), once every 2wk, for a total of 4 times. The patients in both groups were followed up at 1wk after drug treatment, before the first IPL treatment(V1), before the third treatment(V2)of IPL, and at 2wk after the fourth treatment(V3)of IPL, respectively. Data including ocular surface disease index(OSDI), the morphology of meibomian margin under the slit lamp, the nature of meibomian gland(MG)secretion, the difficulty of excretion of MG secretion, non-invasive tear film break-up time(NIBUT), red eye index and corneal fluorescent staining were collected. Moreover, the best corrected visual acuity(BCVA), intraocular pressure, anterior segment and fundus were examined to observe the occurrence of complications.RESULTS: There was no statistically significant difference in the indexes of the patients in both groups before treatment, including OSDI, morphology of meibomian margin, nature of MG secretion, the difficulty of excretion of MG secretion, NIBUT, ocular surface hyperemia and corneal fluorescein sodium staining(P>0.05). In V2 and V3, the indexes of the patients in both groups, including OSDI, morphology of meibomian margin, nature of MG secretion, the difficulty of excretion of MG secretion, ocular surface hyperemia and corneal fluorescein sodium staining, were lower than those before treatment(V1), while the NIBUT was longer than that before treatment, the differences were statistically significant(all P<0.05). There was difference between the groups(P<0.05). Those indexes improved more obvious in the treatment group. No obvious complications occurred in all patients.CONCLUSION: IPL comprehensive treatment have better effect on reducing the inflammation of ocular surface, and improving the function of MG, and it can be used as a new option for physical therapy of BKC.

Objective: To investigate the effects of exosomes from human adipose-derived mesenchymal stem cells (ADSCs) on inflammatory response of mouse RAW264.7 cells and wound healing of full-thickness skin defects in mice. Methods: The experimental research methods were adopted. The discarded adipose tissue was collected from 3 female patients (aged 10-25 years) who underwent abdominal surgery in the First Affiliated Hospital of Air Force Medical University. ADSCs were extracted from the adipose tissue by collagenase Ⅰ digestion and identified with flow cytometry. Exosomes were extracted from the human ADSCs by differential ultracentrifugation, the morphology of the exosomes was observed by transmission electron microscopy, the particle diameter of the exosomes was detected by nanoparticle tracking analyzer, and the protein expressions of CD9, CD63, tumor susceptibility gene 101 (TSG101), and β-actin were detected by Western blotting. The human ADSCs exosomes (ADSCs-Exos) and RAW264.7 cells were co-cultured for 12 h, and the uptake of RAW264.7 cells for human ADSCs-Exos was observed. The RAW264.7 cells were divided into phosphate buffer solution (PBS) group stimulated with PBS for suitable time, endotoxin/lipopolysaccharide (LPS) stimulation 2 h group, LPS stimulation 4 h group, LPS stimulation 6 h group, LPS stimulation 12 h group, and LPS stimulation 24 h group stimulated with LPS for corresponding time, with 3 wells in each group, and the mRNA expressions of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), IL-6, and IL-10 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) method. The RAW264.7 cells were divided into PBS group, LPS alone group, and LPS+ADSCs-Exos group, with 3 wells in each group, which were dealt correspondingly for the time screened out in the previous experiment, the mRNA expressions of IL-1β, TNF-α, IL-6, IL-10, trasforming growth factor β (TGF-β,) and vascular endothelial growth factor (VEGF) were detected by real time fluorescence quantitative RT-PCR method, and the protein expressions of inducible nitric oxide synthase (iNOS) and arginase 1 (Arg1) were detected by Western blotting. Twenty-four 8-week-old male BALB/c mice were divided into PBS group and ADSCs-Exos group according to the random number table, with 12 mice in each group, and a full-thickness skin defect wound with area of 1 cm×1 cm was inflicted on the back of each mouse. Immediately after injury, the wounds of mice in the two groups were dealt correspondingly. On post injury day (PID) 1, the concentration of IL-1β and TNF-α in serum were detected by enzyme-linked immunosorbent assay, and the mRNA expressions of IL-1β, TNF-α, and IL-6 were detected by real time fluorescence quantitative RT-PCR method. On PID 3, 6, 9, 12, and 15, the wound healing was observed and the wound non-healing rate was calculated. On PID 15, the defect length of skin accessory and collagen volume fraction (CVF) were detected by hematoxylin eosin staining and Masson staining, respectively, the CD31 expression and neovascularization were detected by immunohistochemistry, and the ratio of Ki67 positive cells, the ratio of iNOS and Arg1 double positive cells, and the ratio of iNOS positive cells to Arg1 positive cells and their fluorescence intensities were detected by immunofluorescence method. The number of samples in animal experiments was 6. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and independent sample t test. Results: At 12 h of culture, the cells exhibited a typical spindle shape, which were verified as ADSCs with flow cytometry. The exosomes with a vesicular structure and particle diameters of 29-178 nm, were positively expressed CD9, CD63, and TSG101 and negatively expressed β-actin. After 12 h of co-culture, the human ADSCs-Exos were endocytosed into the cytoplasm by RAW264.7 cells. The mRNA expressions of IL-1β, TNF-α, IL-6, and IL-10 of RAW264.7 cells in LPS stimulation 2 h group, LPS stimulation 4 h group, LPS stimulation 6 h group, LPS stimulation 12 h group, and LPS stimulation 24 h group were significantly higher than those in PBS group (with t) values of 39.10, 14.55, 28.80, 4.74, 48.80, 22.97, 13.25, 36.34, 23.12, 18.71, 29.19, 41.08, 11.68, 18.06, 8.54, 43.45, 62.31, 22.52, 21.51, and 37.13, respectively, P<0.01). The stimulation 12 h with significant expressions of all the inflammatory factors was selected as the time point in the following experiment. After stimulation of 12 h, the mRNA expressions of IL-1β, TNF-α, IL-6, and IL-10 of RAW264.7 cells in LPS alone group were significantly higher than those in PBS group (with t values of 44.20, 51.26, 14.71, and 8.54, respectively, P<0.01); the mRNA expressions of IL-1β, TNF-α, and IL-6 of RAW264.7 cells in LPS+ADSCs-Exos group were significantly lower than those in LPS alone group (with t values of 22.89, 25.51, and 8.03, respectively, P<0.01), while the mRNA expressions of IL-10, TGF-β, and VEGF were significantly higher than those in LPS alone group (with t values of 9.89, 13.12, and 7.14, respectively, P<0.01). After stimulation of 12 h, the protein expression of iNOS of RAW264.7 cells in LPS alone group was significantly higher than that in PBS group and LPS+ADSCs-Exos group, respectively (with t values of 11.20 and 5.06, respectively, P<0.05 or P<0.01), and the protein expression of Arg1 was significantly lower than that in LPS+ADSCs-Exos group (t=15.01, P<0.01). On PID 1, the serum concentrations of IL-1β and TNF-α and the mRNA expressions of IL-1β, TNF-α, and IL-6 in wound tissue of mice in ADSCs-Exos group were significantly those in lower than PBS group (with t values of 15.44, 12.24, 9.24, 7.12, and 10.62, respectively, P<0.01). On PID 3, 6, 9, 12, and 15 d, the wound non-healing rates of mice in ADSCs-Exos group were (73.2±4.1)%, (53.8±3.8)%, (42.1±5.1)%, (24.1±2.8)%, and 0, which were significantly lower than (82.5±3.8)%, (71.2±4.6)%, (52.9±4.1)%, (41.5±3.6)%, and (14.8±2.5)% in PBS group, respectively (with t values of 4.77, 8.93, 5.54, 7.63, and 7.59, respectively, P<0.01). On PID 15, the defect length of skin accessory in wounds of mice in PBS group was significantly longer than that in ADSCs-Exos group (t=9.50, P<0.01), and the CVF was significantly lower than that in ADSCs-Exos group (t=9.15, P<0.01). On PID 15, the CD31 expression and the number of new blood vessels (t=12.99, P<0.01), in wound tissue of mice in ADSCs-Exos group were significantly more than those in PBS group, and the ratio of Ki67 positive cells was significantly higher than that in PBS group (t=7.52, P<0.01). On PID 15, the ratio of iNOS and Arg1 double positive cells in wound tissue of mice in PBS group was (12.33±1.97)%, which was significantly higher than (1.78±0.29)% in ADSCs-Exos group (t=13.04, P<0.01), the ratio of iNOS positive cells and the fluorescence intensity of iNOS were obviously higher than those of ADSCs-Exos group, and the ratio of Arg1 positive cells and the fluorescence intensity of Arg1 were obviously lower than those of ADSCs-Exos group. Conclusions: The human ADSCs-Exos can alleviate inflammatory response of mouse RAW264.7 cells, decrease macrophage infiltration and secretion of the pro-inflammatory cytokines, increase the secretion of anti-inflammatory cytokines to promote neovascularization and cell proliferation in full-thickness skin defect wounds of mice, hence accelerating wound healing.
Animals ; Exosomes ; Female ; Humans ; Male ; Mesenchymal Stem Cells ; Mice ; Skin ; Vascular Endothelial Growth Factor A ; Wound Healing

OBJECTIVE: To explore the characteristics and influencing factors of high-risk factors and adverse outcomes of pregnancy in different occupational populations in a medical college community. METHODS: A total of 719 pregnant women in a medical college community were selected by convenient sampling method and divided into medical staff group(218 women) and non-medical staff group(501 women, including 138 teaching staff subgroup, 129 administrative service staff subgroup and 234 other occupation subgroup). The detection rate of high-risk factors and adverse outcomes of pregnancy were compared among these groups. RESULTS: Among the study subjects with the top five detection rates, high-risk factors of pregnancy were abnormal body mass index, advanced age, diabetes mellitus, scarred uterus and abnormal thyroid function, with the detection rate of 21.4%, 17.5%, 9.7%, 7.5% and 7.5%, respectively. The detection rates of pregnancy high-risk factors≥two, adverse pregnancy outcome, adverse fetal outcome, miscarriage and low birth weight in the medical group were significantly lower than those in the non-medical staff group(all P<0.05). The detection rates of pregnancy risk factors, primary screening risk factors, scarred uterus and pregnancy anemia in the teaching staff subgroup were significantly higher than those in the other occupation subgroups(all P<0.008). The detection rates of pregnancy high-risk factors ≥two and advanced age in the teaching staff subgroup were higher than those in medical staff group(all P<0.008). The detection rates of adverse pregnancy outcome and miscarriage in the administrative service staff subgroup were higher than those in medical staff group(all P<0.008). Multivariate logistic regression analysis results showed that occupation, advanced age, parity and scarred uterus were the main influencing factors of adverse pregnancy outcome(all P<0.05). CONCLUSION: There are differences in pregnancy high-risk factors and adverse pregnancy outcomes among different occupational groups. The teaching staff subgroup has a relatively higher detection rate of pregnancy high-risk factors, and the adverse pregnancy outcomes and miscarriage were relatively higher in the administrative service staff subgroup, compared with the medical staff group.

【Objective】 To investigate the negative emotions among university students during home quarantine under the epidemic of coronavirus disease 2019 (COVID-19) so as to provide scientific basis for psychological counseling. 【Methods】 We conducted an online questionnaire survey to collect 405 undergraduates’ and postgraduates’ DASS-21 scale scores and responses to epidemic prevention knowledge, and analyzed the university students’ negative emotions during the epidemic and influencing factors. 【Results】 Of the 405 students surveyed, 178 individuals (44.0%) had depression, 171(42.2%) felt anxious, and 119(29.4%) felt stressed. Individuals in the DASS21+ group spent much time on entertainment on the cell phone daily than those in the DASS21- group (P<0.001), and the degree of impact by COVID-19 on daily life was significantly higher (P<0.001). Correlation analysis showed that entertainment time on the cell phone, the severity of COVID-19, and the impact of the epidemic were positively related to DASS-21 scores, with correlation coefficients of 0.231, 0.143 and 0.259, respectively (all P<0.01). Binary logistic regression analysis found that mobile entertainment time for over 5 hours per day (OR=3.370, 95% CI: 1.369-8.294, P=0.008) was a risk factor for negative emotions during quarantine at home. 【Conclusion】 During quarantine at home in the epidemic of COVID-19, nearly half of university students are prone to breed negative emotions, like depression, anxiety or stress, which may be related to long-term mobile phone entertainment.

As a serine hydrolase, monoacylglycerol lipase (MAGL) is principally responsible for the metabolism of 2-arachidonoylglycerol (2-AG) in the central nervous system (CNS), leading to the formation of arachidonic acid (AA). Dysfunction of MAGL has been associated with multiple CNS disorders and symptoms, including neuroinflammation, cognitive impairment, epileptogenesis, nociception and neurodegenerative diseases. Inhibition of MAGL provides a promising therapeutic direction for the treatment of these conditions, and a MAGL positron emission tomography (PET) probe would greatly facilitate preclinical and clinical development of MAGL inhibitors. Herein, we design and synthesize a small library of fluoropyridyl-containing MAGL inhibitor candidates. Pharmacological evaluation of these candidates by activity-based protein profiling identified