BACKGROUND:Macrophage polarization plays a key role in chronic inflammatory joint diseases such as rheumatoid arthritis.Cerium dioxide(CeO2)nanoparticles have a wide range of biomedical applications such as modulating the local inflammatory microenvironment of tissues.OBJECTIVE:To investigate the role of CeO2 nanoparticles on macrophage polarization and inflammatory factor expression,as well as inflammatory modulation in a co-culture system of macrophages and fibroblasts.METHODS:(1)CeO2 nanoparticles were dispersed and observed morphologically by transmission electron microscopy.(2)Human leukemia monocytes(THP-1)were induced to differentiate and establish the M1 macrophage pro-inflammatory cell model of rheumatoid arthritis.The cells were divided into M0 group(undifferentiated macrophages),M1 group(successful macrophage modeling),CeO2 nanoparticle treatment group(M1 group with CeO2 nanoparticle treatment),and dexamethasone control group(M1 group with dexamethasone treatment)and incubated for 48 hours.The effects of CeO2 nanoparticles on the expression of inflammatory factors(endogenous nitric oxide synthase,CD86,CD80)in M1 macrophages and M1 macrophage phenotype(CD80,CD206)were detected by RT-qPCR,western blot assay,and flow cytometry.(3)A co-culture system of macrophages and fibroblasts was established,and CeO2 nanoparticles acted on the upper macrophages.The regulation of CeO2 nanoparticles on the expression of inflammatory factors(interleukin-6,tumor necrosis factor-α,cyclooxygenase-2,and endogenous nitric oxide synthase)of fibroblasts in the co-culture system was observed at the mRNA and protein levels.RESULTS AND CONCLUSION:(1)Transmission electron microscopy showed that the diameter of CeO2 nanoparticles was(19.5±2.0)nm.(2)Compared with the M0 group,the mRNA of endogenous nitric oxide synthase and CD86,and the protein expression of endogenous nitric oxide synthase and CD80 in the M1 group were upregulated.Compared with the M1 group,the mRNA expression of endogenous nitric oxide synthase and CD86,and the protein expression of endogenous nitric oxide synthase and CD80 in the CeO2 nanoparticle treatment group were downregulated.Flow cytometry showed that 20 nm CeO2 nanoparticles downregulated the number of M1 macrophages.(3)Compared with the M1 group,20 nm CeO2 nanoparticles downregulated the mRNA and protein expression of inflammatory factors(tumor necrosis factor α,interleukin 6,cyclooxygenase 2,and endogenous nitric oxide synthase)in the co-culture system HFL1 cells.(4)The results showed that 20 nm CeO2 nanoparticles can alleviate inflammation in the co-culture system by inhibiting the expression of pro-inflammatory factors in M1 macrophages,providing a new idea for the treatment of inflammatory diseases such as rheumatoid arthritis.

BACKGROUND:Macrophage polarization plays a key role in chronic inflammatory joint diseases such as rheumatoid arthritis.Cerium dioxide(CeO2)nanoparticles have a wide range of biomedical applications such as modulating the local inflammatory microenvironment of tissues.OBJECTIVE:To investigate the role of CeO2 nanoparticles on macrophage polarization and inflammatory factor expression,as well as inflammatory modulation in a co-culture system of macrophages and fibroblasts.METHODS:(1)CeO2 nanoparticles were dispersed and observed morphologically by transmission electron microscopy.(2)Human leukemia monocytes(THP-1)were induced to differentiate and establish the M1 macrophage pro-inflammatory cell model of rheumatoid arthritis.The cells were divided into M0 group(undifferentiated macrophages),M1 group(successful macrophage modeling),CeO2 nanoparticle treatment group(M1 group with CeO2 nanoparticle treatment),and dexamethasone control group(M1 group with dexamethasone treatment)and incubated for 48 hours.The effects of CeO2 nanoparticles on the expression of inflammatory factors(endogenous nitric oxide synthase,CD86,CD80)in M1 macrophages and M1 macrophage phenotype(CD80,CD206)were detected by RT-qPCR,western blot assay,and flow cytometry.(3)A co-culture system of macrophages and fibroblasts was established,and CeO2 nanoparticles acted on the upper macrophages.The regulation of CeO2 nanoparticles on the expression of inflammatory factors(interleukin-6,tumor necrosis factor-α,cyclooxygenase-2,and endogenous nitric oxide synthase)of fibroblasts in the co-culture system was observed at the mRNA and protein levels.RESULTS AND CONCLUSION:(1)Transmission electron microscopy showed that the diameter of CeO2 nanoparticles was(19.5±2.0)nm.(2)Compared with the M0 group,the mRNA of endogenous nitric oxide synthase and CD86,and the protein expression of endogenous nitric oxide synthase and CD80 in the M1 group were upregulated.Compared with the M1 group,the mRNA expression of endogenous nitric oxide synthase and CD86,and the protein expression of endogenous nitric oxide synthase and CD80 in the CeO2 nanoparticle treatment group were downregulated.Flow cytometry showed that 20 nm CeO2 nanoparticles downregulated the number of M1 macrophages.(3)Compared with the M1 group,20 nm CeO2 nanoparticles downregulated the mRNA and protein expression of inflammatory factors(tumor necrosis factor α,interleukin 6,cyclooxygenase 2,and endogenous nitric oxide synthase)in the co-culture system HFL1 cells.(4)The results showed that 20 nm CeO2 nanoparticles can alleviate inflammation in the co-culture system by inhibiting the expression of pro-inflammatory factors in M1 macrophages,providing a new idea for the treatment of inflammatory diseases such as rheumatoid arthritis.

BACKGROUND: Osteoarthritis (OA) is a prevalent joint disorder that significantly impairs quality of life among elderly individuals because of chronic pain and physical disability. As the global burden of OA continues to rise, novel therapeutic strategies are urgently needed. Kaempferide (KA), a flavonoid derived from traditional Chinese herbal medicine, is known for its anti-inflammatory properties. However, the effect of KA on the progression of OA has not been well investigated. This study aimed to explore the therapeutic potential of KA in an OA model and investigate the underlying mechanisms via transcriptomic sequencing. METHODS: An in vitro OA model was established using SW1353 cells treated with interleukin-1 beta (IL-1β) and different concentrations of KA (30, 60, or 90 μmol/L) for 24 h. The anti-inflammatory effects of KA were assessed using quantitative real-time polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), and Western blotting. In vivo , a papain-induced OA rat model was used to evaluate the therapeutic effects of KA through histological and behavioral analyses. Transcriptomic sequencing was performed to explore the differentially expressed genes (DEGs) and related signaling pathways. Statistical analysis was conducted using one-way analysis of variance. RESULTS: KA significantly increased cell viability in the OA chondrocyte model and downregulated the expression of inflammatory cytokines and cartilage degradation markers, with the greatest reduction observed at 90 μmol/L. In vivo , KA treatment mitigated cartilage degradation and improved gait behavior in OA rats. Transcriptomic analysis revealed substantial modulation of DEGs, implicating the hypoxia-inducible factor-1 (HIF-1) signaling pathway as a key mechanism. Further blocking and rescue experiments revealed that KA regulated key molecules within the HIF-1 pathway, specifically interferon-gamma (IFN-γ) and hypoxia-inducible factor 1-alpha (HIF-1α), confirming their critical roles in mediating the therapeutic effects of KA. CONCLUSION KA inhibited the progression of OA by targeting the HIF-1 signaling pathway, reducing inflammation, and cartilage degradation.
Animals ; Osteoarthritis/metabolism* ; Signal Transduction/drug effects* ; Rats ; Rats, Sprague-Dawley ; Humans ; Male ; Hypoxia-Inducible Factor 1/metabolism* ; Interleukin-1beta

Objective:To evaluate the feasibility of utilizing artificial intelligence (AI) to assist junior anesthesia residents in making the preoperative anesthesia plans.Methods:Forty anesthesia residents in their third year of training, who had obtained their practicing physician qualifications in the Yangzhou area, were assigned into 4 groups ( n=10 each) using a random number table method: Chat-GPT combined with Bing chat group (C-G-B group), Chat-GPT group (C-G group), Bing chat group (B group), and control group (C group). Fifty patients undergoing elective non-cardiac surgery were selected from the anesthesia clinic as teaching cases. C-G-B, C-G and B groups utilized different AI tools to assist trainees in designing anesthesia plans, producing standardized textual outputs. Each trainee underwent a baseline knowledge test through a professional theory examination prior to enrollment. The completeness and accuracy of the preoperative anesthesia plans were evaluated and scored by 3 chief anesthesiologists. The total time spent on plan formulation and satisfaction scores regarding AI tool feedback were recorded. An analysis was conducted based on the American Society of Anesthesiologists (ASA) Physical Status classification of teaching cases. Results:In ASA Physical Status classification Ⅰ and Ⅱ teaching cases, there was no statistically significant difference in completeness and accuracy scores among the four groups ( P>0.05). In ASA Physical Status classification Ⅲ teaching cases, compared to C group, the completeness and accuracy scores were significantly increased in C-G-B, C-G and B groups, with the highest scores observed in C-G-B group ( P<0.05). Among all teaching cases (ASA Physical Status classification Ⅰ-Ⅲ), the total time spent was significantly shortened in C-G and B groups as compared to C and C-G-B groups ( P<0.05). There was no statistically significant difference in the total time spent between C-G group and C-G-B group ( P>0.05). Compared to C-G and B groups, the satisfaction score was significantly decreased in C-G-B group ( P<0.05). Conclusions:For ASA Physical Status classification Ⅲ patients, using AI to assist junior anesthesia residents in making preoperative anesthesia plans may offer advantages. Although combining the use of Chat-GPT and Bing chat can further improve the completeness and accuracy of anesthesia plan development, it may require more time.

Objective:To evaluate the relationship between silent information regulator 1 (SIRT1) and ferroptosis during curcumin-induced reduction of acute lung injury in a mouse model of sepsis.Methods:One hundred and fifty-two SPF-grade male C57BL/6J mice, aged 8 weeks, weighing 23-27 g, were divided into 4 groups ( n=38 each) using a random number table method: sham operation group (C group), sepsis group (S group), curcumin group (Cur group) and curcumin plus SIRT1 inhibitor EX527 group (CE group). Curcumin 200 mg/kg was administered by intragastric gavage every day in Cur group. Curcumin 200 mg/kg was administered by intragastric gavage every day and EX527 5 mg/kg was intraperitoneally injected in CE group. The equal volume of solvent dimethyl sulfoxide (DMSO) was given in C group and S group. Sepsis model was developed by cecal ligation and puncture (CLP) after 5 days of consecutive administration in anesthetized animals. Twenty mice in each group were randomly selected to observe the survival condition within 7 days after CLP. The bronchoalveolar lavage fluid (BALF) was collected at 24 h after developing the model to determine the concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), IL-6 and IL-18 (by enzyme-linked immunosorbent assay), and the lung tissues were obtained for microscopic examination of the pathological changes which were scored and for determination of wet-to-dry lung weight (W/D) ratio, contents of glutathione (GSH), malondialdehyde (MDA) and iron (by colorimetry), and expression of SIRT1, glutathione peroxidase 4 (GPX4) and Acyl-CoA synthetase long chain family member 4 (ACSL4) (by Western blot). Results:Compared with C group, the 7-day survival rate after CLP was significantly decreased, the concentrations of TNF-α, IL-1β, IL-6 and IL-18 in BALF, W/D ratio and lung injury score were increased, the content of GSH in lung tissues was decreased, the contents of MDA and iron were increased, the expression of SIRT1 and GPX4 was down-regulated, and the expression of ACSL4 was up-regulated in S group ( P<0.05). Compared with S group, the 7-day survival rate after CLP was significantly increased, the concentrations of TNF-α, IL-1β, IL-6 and IL-18 in BALF, W/D ratio and lung injury score were decreased, the content of GSH was increased, the contents of MDA and iron were decreased, the expression of SIRT1 and GPX4 was up-regulated, and the expression of ACSL4 was down-regulated in Cur group ( P<0.05). Compared with Cur group, the 7-day survival rate after CLP was significantly decreased, the concentrations of TNF-α, IL-1β, IL-6 and IL-18 in BALF, W/D ratio and lung injury score were increased, the content of GSH was decreased, the contents of MDA and iron were increased, the expression of SIRT1 and GPX4 was down-regulated, and the expression of ACSL4 was up-regulated in CE group ( P<0.05). Conclusions:The mechanism by which curcumin attenuates acute lung injury may be related to activation of SIRT1 and further inhibition of ferroptosis in mice.

Objective To observe in vitro the effect of mechanical stress at different intensities on the pro-liferation, apoptosis and extracellular matrix of degenerative human nucleus pulposus cells. Methods The cells were isolated and cultured in vitro, and divided into a control group, a low-intensity group, a medium-intensity group and a high-intensity group. The low-, medium- and high-intensity groups were stretched mechanically by 1000 μ, 2000μor 4000μrespectively for 6 hours using a four-point bending system, while the control group was not stressed. Flow cytometry was used to explore any changes in the cell cycle and the proliferation index ( PI) . The expression of proliferating cell nuclear antigen ( PCNA) , B-cell lymphoma-2 ( BCL-2)/Bax, collagen II and aggrecan were meas-ured using real-time quantitative polymerase chain reactions. Results The mechanical stretching significantly influ-enced proliferation, apoptosis and the extracellular matrices compared with the control group. The PI and PCNA ex-pression increased at first and then decreased gradually with the exercise intensity. Compared with the control group, the mRNA expression level of Bcl-2/Bax increased significantly to 1.53 times that of the control group after 1000 μstretching, but to only 0.71 times that of the control group at 2000μand 0.43 times at 4000μ. The gene expression of collagen II increased significantly by 1. 1 times and that of aggrecan by 1. 3 times after 1000 μ stress stimulation compared with the control group ( P≤0.05) . However, the expression of collagen II and aggrecan was inhibited sig-nificantly at 2000 and 4000 μ , with the lowest levels at 4000 μ (P≤0.05). Conclusion Stretching at different intensities has different effects on the proliferation, apoptosis and extracellular matrix of human pulposus cells with degenerate nuclei.

Objective To evaluate the effect of ventilation with different positive end-expiratory pressures (PEEPs) on intracranial pressure in the patients undergoing gynecological laparoscopic surgery.Methods Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients,aged 25-55 yr,with body mass index of 18-27 kg/m2,scheduled for elective gynecological endoscopic surgery,were divided into 3 groups (n=20 each) using a random number table method:routine ventilation group (group A),4 cmH2O PEEP group (group B) and 8 cmH2O PEEP group (group C).The patients were mechanically ventilated with PEEP 4 or 8 cmH2O at 5 min of head-down tilt after start of pneumoperitoneum in group B and group C.The patients were mechanically ventilated in volume-controlled mode,with tidal volume 7 ml/kg,inspired oxygen concentration 50％ and inspiratory/expiratory ratio 1 ∶ 2.Blood samples were collected from the radial artery for measurement of PaCO2 and PaO2 at 5 min after tracheal intubation (T0),5,15 and 30 min of head-down tilt (T1-3),and 5 min of the supine position after the end of pneumoperitoneum (T4).The peak airway pressure (Ppeak) was recorded,dynamic pulmonary compliance (Cdyn)was calculated,and optic nerve sheath diameter was measured using an ultrasonic apparatus at T0-4.Cognitive function was assessed at 1 day before surgery and 7 days after surgery using Mini-Mental State Examination.Results Compared with group A,Ppeak was significantly increased at T1-4,and PaO2 was increased at T2 in group B,and Ppeak and PaO2 were increased at T1-4,and Cdyn was increased at T1,2 in group C (P＜0.05).Compared with group B,Ppeak was significantly increased at T4,and Cdyn was increased at T2 in group C (P＜0.05).There was no significant difference in optic nerve sheath diameter or Mini-Mental State Examination score at each time point among three groups (P＞0.05).No patients developed cognitive dysfunction at 7 days after surgery in three groups.Conclusion Ventilation with different PEEPs causes no increase in intracranial pressure of the patients undergoing gynecological laparoscopic surgery.