BACKGROUND:Pinoresinol diglucoside promotes bone formation and bone matrix synthesis and accelerates bone tissue repair.However,the mechanism of action and effects of this compound in osteoblasts need to be further explored. OBJECTIVE:To investigate the effect and mechanism of action of pinoresinol diglucoside on dexamethasone-treated osteoblasts based on the Wnt/β-catenin signaling pathway. METHODS:Different concentrations of dexamethasone groups and pinoresinol diglucoside groups were set to treat osteoblasts for 24 hours,and the optimal intervention concentrations were screened.Osteoblasts were treated with dexamethasone,pinoresinol diglucoside and inhibitor XAV-939.Then,control group,dexamethasone group,XVA-939 group,pinoresinol diglucoside group,pinoresinol diglucoside+XVA-939 group were set up.Cell counting kit-8 assay was used to detect cell activity.Alkaline phosphatase activity and caspase3/7 enzyme activity in cells were detected.Annexin V/PI staining and EdU assay were used to detect cell apoptosis and proliferation.Real-time qPCR and western blot were used to detect the mRNA and protein expression levels of Wnt3a,β-catenin,c-myc,osteocalcin,and type I collagen,respectively. RESULTS AND CONCLUSION:After dexamethasone and pinoresinol diglucoside intervened in osteoblasts for 24 hours,10 μmol/L dexamethasone was found to be the optimal intervention concentration for cell inhibition,and cell proliferation was most pronounced at a concentration of pinoresinol diglucoside of 100 μmol/L.Compared with the dexamethasone group,alkaline phosphatase activity was significantly enhanced(P<0.05)and caspase3/7 enzyme activity was significantly reduced(P<0.05)in the pinoresinol diglucoside group.Annexin V/PI staining and cell proliferation assay by EdU method showed that pinoresinol diglucoside inhibited apoptosis and promoted proliferation of osteoblasts after dexamethasone intervention.The mRNA and protein expression levels of Wnt3a,β-catenin,c-myc,osteocalcin,and type I collagen were significantly higher in the pinoresinol diglucoside group and pinoresinol diglucoside+XVA-939 group compared with the dexamethasone and XVA-939 groups(P<0.05).To conclude,pinoresinol diglucoside can inhibit osteoblast apoptosis after dexamethasone intervention,protect osteoblast activity and promote osteoblast proliferation by activating the Wnt/β-catenin signaling pathway,which may play a role in delaying steroid-induced osteonecrosis of the femoral head.

Objective To analyze the diversity and composition of gut microbiota in individuals with circadian rhythm disruption and their correlation with cognition.Methods Night shift workers and regular shift workers were subjected from our hospital during August 2022 and October 2024.The participants with circadian rhythm disorders were assigned into an experimental group(n=24),and those with normal circadian rhythms were into a control group(n=24).Their height,weight,age,gender,body mass index(BMI)and fresh fecal samples were collected,and Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)were used to evaluate their mental status.Metagenomics,Alpha and Beta diversity analyses,Linear Discriminant Analysis Effect Size(LEfSe),and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were employed to investigate the diversity and function characteristics of gut microbiota in the participants.Results There were no statistical differences between the 2 groups in baseline data,such as height,weight,gender,age,and BMI(P>0.05).Alpha diversity analysis indicated that no statistical differences were observed in the ACE,Chao1,Shannon,or Simpson indices between the 2 groups,while beta diversity analysis revealed significant differences(P<0.01),suggesting different structure of gut microbiota between them.In the experimental group,the abundance of Faecalibacterium prausnitzii and Agathobacter rectalis was decreased,while that of Escherichia coli and Phocaeicola vulgaratus was increased,with significant differences when compared with the control group(P<0.05).Additionally,KEGG functional analysis showed that the experimental group had obviously higher expression levels in Th17 cell differentiation and the IL-17 signaling pathway than the control group(P<0.05).Agathobacter rectalis and Faecalibacterium prausnitzii were positively correlated with MoCA score and MMSE score(P<0.05,P<0.01).Agathobacter rectalis was negatively correlated with the IL-17 signaling pathway and Th17 cell differentiation.Conclusion Individuals with circadian rhythm disorders have significant changes in the structure and function of gut microbiota when compared to those with normal circadian rhythms.Agathobacter rectalis may be involved in the regulation of the IL-17 signaling pathway and differentiation of Th17 cells,thereby possibly impacting the increases of cognitive score related to circadian rhythm disorders.

Objective:To investigate the safety of early antiplatelet therapy for non-cardioembolic mild stroke patients with thrombocytopenia.Methods:Data of acute ischemic stroke patients with baseline National Institutes of Health Stroke Scale(NIHSS)score≤3 and a platelet count<100×109/L were obtained from a multicenter register.Those who required anticoagulation or had other contraindications to antiplatelet therapy were excluded.Short-term safety outcomes were in-hospital bleeding events,while the long-term safety outcome was a 1-year all-cause death.The short-term neurological outcomes were evaluated by modified Rankin scale(mRS)score at discharge.Results:A total of 1868 non-cardioembolic mild stroke patients with thrombocytopenia were enrolled.Multivariate regression analyses showed that mono-antiplatelet therapy significantly increased the proportion of mRS score of 0-1 at discharge(OR=1.657,95%CI:1.253-2.192,P<0.01)and did not increase the risk of intracranial hemorrhage(OR=2.359,95%CI:0.301-18.503,P>0.05),compared with those without antiplatelet therapy.However,dual-antiplatelet therapy did not bring more neurological benefits(OR=0.923,95%CI:0.690-1.234,P>0.05),but increased the risk of gastrointestinal bleeding(OR= 2.837,95%CI:1.311-6.136,P<0.01)compared with those with mono-antiplatelet therapy.For patients with platelet counts≤75×109/L and>90×109/L,antiplatelet therapy significantly improved neurological functional outcomes(both P<0.05).For those with platelet counts(>75-90)×109/L,antiplatelet therapy resulted in a significant improvement of 1-year survival(P<0.05).For patients even with concurrent coagulation abnormalities,mono-antiplatelet therapy did not increase the risk of various types of bleeding(all P>0.05)but improved neurological functional outcomes(all P<0.01).There was no significant difference in the occurrence of bleeding events,1-year all-cause mortality risk,and neurological functional outcomes between aspirin and clopidogrel(all P>0.05).Conclusions:For non-cardioembolic mild stroke patients with thrombocytopenia,antiplatelet therapy remains a reasonable choice.Mono-antiplatelet therapy has the same efficiency as dual-antiplatelet therapy in neurological outcome improvement with lower risk of gastrointestinal bleeding.

Diabetic wound is a complication of diabetes, which is difficult to heal and easy to turn into chronic wound. Compared with ordinary wound healing, diabetic wounds stay in the inflammatory stage and can not enter the proliferative stage. This is because the proportion of pro-inflammatory cytokines increases, leading to the imbalance of inflammatory cells and the accumulation of a large number of inflammatory factors. The production and release of inflammatory factors is closely related to pyroptosis. Pyroptosis is a kind of programmed death mode of inflammatory cells. Mediated by inflammasome, pyroptosis is transduced through typical and atypical inflammasome signaling pathways, resulting in the formation of pores on the cell membrane and inducing cell death. In this process, a large number of pro-inflammatory cytokines are released to maintain the inflammatory environment of wounds, hindering the development of diabetic wounds to the proliferative stage and remodeling stage, thus inhibiting the healing of diabetic wounds. This article reviews the mechanism of pyroptosis and its effect on diabetic wounds, the potential significance of inhibiting pyroptosis in the treatment of diabetic wounds, and the existing problems.

Background::Triple-negative breast cancer (TNBC) is a type of highly invasive breast cancer with a poor prognosis. According to new research, long noncoding RNAs (lncRNAs) play a significant role in the progression of cancer. Although the role of lncRNAs in breast cancer has been well reported, few studies have focused on TNBC. This study aimed to explore the biological function and clinical significance of forkhead box C1 promoter upstream transcript (FOXCUT) in triple-negative breast cancer.Methods::Based on a bioinformatic analysis of the cancer genome atlas (TCGA) database, we detected that the lncRNA FOXCUT was overexpressed in TNBC tissues, which was further validated in an external cohort of tissues from the General Surgery Department of the First Affiliated Hospital of Nanjing Medical University. The functions of FOXCUT in proliferation, migration, and invasion were detected in vitro or in vivo. Luciferase assays and RNA immunoprecipitation (RIP) were performed to reveal that FOXCUT acted as a competitive endogenous RNA (ceRNA) for the microRNA miR-24-3p and consequently inhibited the degradation of p38. Results::lncRNA FOXCUT was markedly highly expressed in breast cancer, which was associated with poor prognosis in some cases. Knockdown of FOXCUT significantly inhibited cancer growth and metastasis in vitro or in vivo. Mechanistically, FOXCUT competitively bounded to miR-24-3p to prevent the degradation of p38, which might act as an oncogene in breast cancer. Conclusion::Collectively, this research revealed a novel FOXCUT/miR-24-3p/p38 axis that affected breast cancer progression and suggested that the lncRNA FOXCUT could be a diagnostic marker and therapeutic target for breast cancer.

Objective:To evaluate the role of NOD-like receptor 3 (NLRP3) inflammasome activation-mediated macrophage polarization in myocardial injury after ischemic stroke in diabetic mice.Methods:Wild-type C57BL/6J mice and NLRP3 -/- mice, aged 4-6 weeks, were fed a high fat diet combined with streptozotocin administration to develop the diabetic model. Twenty-four diabetic wild type C57BL/6J mice and 23 diabetic NLRP3 -/- mice were divided into wild type sham operation group (WT D-SHAM group, n=9) , wild type ischemic stroke group (WT D-MCAO group, n=15) , NLRP3 -/- sham operation group (NLRP3 -/-D-SHAM group, n=9) and NLRP3 -/- ischemic stroke group (NLRP3 -/-D-MCAO group, n=14). The ischemic stroke model was developed by middle cerebral artery occlusion in the animals anesthetized with isoflurane. Echocardiography and electrocardiography were carried out at 3, 7, 14 and 28 days after developing the model. Mice were sacrificed under deep anesthesia, and myocardial tissues were taken at 28 days after surgery for determination of the expression of macrophage marker F4/80 and M2 type macrophage marker CD206 mRNA (by real-time fluorescence quantitative polymerase chain reaction). Results:Compared with WT D-SHAM group, the cardiac output, mass of left ventricle and corrected mass of left ventricle were significantly decreased at 28 days after surgery, and QT interval and QTc interval were prolonged at 14 and 28 days after developing the model in WT D-MCAO group ( P<0.05). Compared with NLRP3 -/-D-SHAM group, the cardiac output, mass of left ventricle and corrected mass of left ventricle were significantly decreased, and QT interval and QTc interval were prolonged at 3 days after surgery in NLRP3 -/-D-MCAO group ( P<0.05). There was no significant difference in CD206 and F4/80 mRNA expression between WT D-SHAM group and WT D-MCAO group and between NLRP3 -/-D-SHAM group and NLRP3 -/-D-MCAO group ( P>0.05). Compared with WT D-MCAO group, the QT interval and QTC interval were significantly shortened at 14 and 28 days after developing the model, and the expression of F4/80 mRNA was down-regulated and the expression of CD206 mRNA was up-regulated at 28 days after developing the model in NLRP3 -/-D-MCAO group ( P<0.05). Conclusions:NLRP3 inflammasome activation-mediated polarization of macrophages to M2 phenotype is involved in myocardial injury after ischemic stroke in diabetic mice.

Gut microbiota dysbiosis is an avenue for the promotion of atherosclerosis (AS) and this effect is mediated partly via the circulating microbial metabolites. More microbial metabolites related to AS vascular inflammation, and the mechanisms involved need to be clarified urgently. Paeonol (Pae) is an active compound isolated from Paeonia suffruticoas Andr. with anti-AS inflammation effect. However, considering the low oral bioavailability of Pae, it is worth exploring the mechanism by which Pae reduces the harmful metabolites of the gut microbiota to alleviate AS. In this study, ApoE^-/- mice were fed a high-fat diet (HFD) to establish an AS model. AS mice were administrated with Pae (200 or 400 mg·kg^-1) by oral gavage and fecal microbiota transplantation (FMT) was conducted. 16S rDNA sequencing was performed to investigate the composition of the gut microbiota, while metabolomics analysis was used to identify the metabolites in serum and cecal contents. The results indicated that Pae significantly improved AS by regulating gut microbiota composition and microbiota metabolic profile in AS mice. We also identified α-hydroxyisobutyric acid (HIBA) as a harmful microbial metabolite reduced by Pae. HIBA supplementation in drinking water promoted AS inflammation in AS mice. Furthermore, vascular endothelial cells (VECs) were cultured and stimulated by HIBA. We verified that HIBA stimulation increased intracellular ROS levels, thereby inducing VEC inflammation via the TXNIP/NLRP3 pathway. In sum, Pae reduces the production of the microbial metabolite HIBA, thus alleviating the ROS/TXNIP/NLRP3 pathway-mediated endothelial inflammation in AS. Our study innovatively confirms the mechanism by which Pae reduces the harmful metabolites of gut microbiota to alleviate AS and proposes HIBA as a potential biomarker for AS clinical judgment.
Animals ; Mice ; Atherosclerosis/drug therapy* ; Diet, High-Fat ; Endothelial Cells ; Inflammation/drug therapy* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Reactive Oxygen Species

Objective:To investigate cognitive function changes and their influential factors in patients with ischemic stroke and leukoaraiosis.Methods:A total of 500 patients with ischemic stroke who received treatment in Yiwu Central Hospital from January 2018 to October 2019 were included in this study. They were divided into simple ischemic stroke group ( n = 200) and ischemic stroke complicated by leukoaraiosis group (combination group, n = 300). The infarct location and the degree of leukoaraiosis in the combination group were analyzed. An additional 150 volunteers who concurrently underwent the Cognitive Function Test in the same hospital were selected as controls. Cognitive function was evaluated using the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA). Patients in the combination group were divided into cognitive impairment group (MoCA score ≥ 26 points) and non-cognitive impairment group (MoCA score < 26 points) according to MoCA score. The risk factors of cognitive impairment in patients with ischemic stroke and leukoaraiosis were analyzed. Results:The scores of the MMSE, MoCA, Clock Drawing Test (CDT), Verbal Fluency Test (VFT), and Digit Span Test (DST) in the control group were (28.93 ± 2.70) points, (28.35 ± 2.74) points, (4.69 ± 1.14) points, (4.94 ± 0.42) points, and (14.33 ± 1.66) points respectively. They were (26.92 ± 2.18) points, (25.02 ± 3.52) points, (3.61 ± 1.60) points, (4.77 ± 0.46) points, and (11.73 ± 1.16) points, respectively in the simple ischemic stroke group and (24.91 ± 2.79) points, (20.70 ± 3.06) points, (2.87 ± 1.23) points, (4.07 ± 0.85) points, and (10.82 ± 0.93) points respectively in the combination group. There were significant differences in the scores of the MMSE, MoCA, CDT, VFT, and DST among the three groups ( F = 124.50, 318.50, 93.43, 112.60, 428.60, all P < 0.001). Significant differences in the scores of the MMSE, MoCA, CDT, VFT, and DST were observed between patients with different degrees of leukoaraiosis ( F = 69.09, 102.40, 20.98, 60.90, 57.00, all P < 0.001). Spearman correlation analysis results showed that the scores of the MMSE, MoCA, CDT, VFT, and DST were negatively correlated with the degree of leukoaraiosis ( r = -0.61, -0.69, -0.43, -0.56, -0.44, all P < 0.05). Logistic regression analysis results showed that age, history of smoking and drinking, history of diabetes, history of stroke, and infarct location were the independent risk factors for cognitive impairment in patients with ischemic stroke and leukoaraiosis. Education level was a protective factor against ischemic stroke and leukoaraiosis. Conclusion:The degree of cognitive impairment in patients with ischemic stroke and leukoaraiosis is related to the degree of leukoaraiosis. Age, history of smoking and drinking, history of diabetes, history of stroke, infarction location, and education level are the influential factors of cognitive impairment.

Objective To investigate the prevalence of musculoskeletal disorders (MSD) in dentists and to analyze the risk factors of MSD to provide suggestions for preventing and reducing MSD in dentists. Methods Through stratified cluster random sampling, 373 dentists were selected from one Hospital of Stomatolagy and five general hospitals among Three-A hospitals in Wuhan as the research objects. The prevalence of MSD was surveyed using the Nordic musculoskeletal disorders standard questionnaire (NMQ) and Numerical Rating Scale (NRS) of pain measurement, and the risk factors of MSD was analyzed with binary Logistic regression. Results The total MSD annual prevalence rate of dentists was 93.3%, with a prevalence rate in neck, shoulder and back, waist and wrist at 78.3%, 70.0%, 56.0% and 36.2% respectively. Through Binary Logistic regression analysis, it was found that the risk factors associated with neck MSD were work fatigue（χ²=24.00，P=0.000）, neck hunching（χ²=23.55，P=0.000）, and shoulder side lift（χ²=24.52，P=0.000）. The risk factors associated with MSD in shoulder and back were work fatigue（χ²=34.64，P=0.000）, neck twisting（χ²=21.68，P=0.000） and wrist bending（χ²=45.87，P=0.000）. The risk factors associated with waist MSD were age（χ²=29.83，P=0.000）, majors（χ²=16.68，P=0.028）, work fatigue（χ²=21.08，P=0.000）, waist bending（χ²=22.88，P=0.000）. The risk factors associated with wrist MSD were gender（χ²=4.17，P=0.041）, majors（χ²=23.47，P=0.001）, working years（χ²=11.63，P=0.009）, physical activities（χ²=9.14，P=0.028）, number of patient visits per day（χ²=18.41，P=0.000）, bending and twisting（χ²=24.12，P=0.000）, wrist twisting（χ²=34.41，P=0.000）, and prevalence of microscope using（χ²=12.09，P=0.020）, while physical exercise was a protective factor for wrist MSD.The differences of the above-mentioned risk factors were statistically significant (P<0.05). Conclusion The prevalence of MSD in dentists is relatively high, and hospital management should strengthen organizational training to help dentists to realize the importance of adopting correct operating posture, a 5-minute breaks during work, prevent MSD at an early stage.