Objective:To assess the effect of joint exposure to multiple air pollutants on sleep structure in patients with stable chronic obstructive pulmonary disease (COPD), identify key air pollutants, and analyze potential influencing factors.Methods:In this panel study, 92 stable COPD patients were recruited. From March 2021 to September 2023 in Beijing, all participants completed 254 nights of sleep monitoring. The total sleep duration, light sleep duration, deep sleep duration and rapid eye movement sleep duration and their respective proportions in total sleep duration were recorded. The exposure levels of fine particulate matter (PM 2.5), inhalable particulate matter (PM 10), nitrogen dioxide (NO 2), ozone (O 3), sulfur dioxide (SO 2), and carbon monoxide (CO) were estimated based on the infiltration factor method and time-activity logs of participants. To assess the lag effect of air pollutants, moving average concentrations of air pollutants from 0-1 day to 0-3 months were calculated. The linear mixed-effect model and Bayesian kernel machine regression (BKMR) model were used to assess the single and joint effects of air pollutants on sleep structure parameters in COPD patients, respectively. Results:All six types of air pollutants were associated with changes in sleep structure, manifesting as an increase in total sleep duration and light sleep proportion and a reduction in deep sleep proportion. The effects of O 3 were strongest at lag 0-6 days, while other air pollutants were at lag 0-3 months. Joint exposure to multiple air pollutants exerted significant joint effects on sleep structure, and NO 2 was identified as the dominant pollutant. NO 2 had a posterior inclusion probability (PIP) greater than 0.5 for light sleep proportion (PIP=0.691) and deep sleep proportion (PIP=0.957). With an interquartile range (IQR) increase of 8.6 μg/m 3 in NO 2 at lag 0-3 months, the light sleep proportion increased by 10.5% (95% CI: 2.2%-19.4%), and the deep sleep proportion decreased by 19.5% (95% CI:-30.6%- -6.8%). Conclusion:Joint exposure to air pollutants is associated with changes in sleep structure in stable COPD patients, and NO 2 may be a key pollutant.

Objective:To assess the effect of joint exposure to multiple air pollutants on sleep structure in patients with stable chronic obstructive pulmonary disease (COPD), identify key air pollutants, and analyze potential influencing factors.Methods:In this panel study, 92 stable COPD patients were recruited. From March 2021 to September 2023 in Beijing, all participants completed 254 nights of sleep monitoring. The total sleep duration, light sleep duration, deep sleep duration and rapid eye movement sleep duration and their respective proportions in total sleep duration were recorded. The exposure levels of fine particulate matter (PM 2.5), inhalable particulate matter (PM 10), nitrogen dioxide (NO 2), ozone (O 3), sulfur dioxide (SO 2), and carbon monoxide (CO) were estimated based on the infiltration factor method and time-activity logs of participants. To assess the lag effect of air pollutants, moving average concentrations of air pollutants from 0-1 day to 0-3 months were calculated. The linear mixed-effect model and Bayesian kernel machine regression (BKMR) model were used to assess the single and joint effects of air pollutants on sleep structure parameters in COPD patients, respectively. Results:All six types of air pollutants were associated with changes in sleep structure, manifesting as an increase in total sleep duration and light sleep proportion and a reduction in deep sleep proportion. The effects of O 3 were strongest at lag 0-6 days, while other air pollutants were at lag 0-3 months. Joint exposure to multiple air pollutants exerted significant joint effects on sleep structure, and NO 2 was identified as the dominant pollutant. NO 2 had a posterior inclusion probability (PIP) greater than 0.5 for light sleep proportion (PIP=0.691) and deep sleep proportion (PIP=0.957). With an interquartile range (IQR) increase of 8.6 μg/m 3 in NO 2 at lag 0-3 months, the light sleep proportion increased by 10.5% (95% CI: 2.2%-19.4%), and the deep sleep proportion decreased by 19.5% (95% CI:-30.6%- -6.8%). Conclusion:Joint exposure to air pollutants is associated with changes in sleep structure in stable COPD patients, and NO 2 may be a key pollutant.

Objective To investigate the release kinetics of Zn2+ from nZCP-loaded polylactic acid/hydroxyapatite(PLA/HA)composite scaffold(PHZ)and determine the optimal nZCP content in the scaffold.Methods The particle size of nZCP was measured by DLS measurement,and PXRD,FTIR,and SEM were used to characterize the scaffolds and nZCP distribution;EDS was used to analyze element composition of the scaffold.Compression strength of the scaffold was determined,and ion release profile was investigated using ICP-MS.The biocompatibility of the materials was evaluated by CCK-8 assay and dead/alive staining of rat bone marrow stem cells(BMSCs)incubated with their aqueous extracts.ALP staining,alizarin red staining,RT-qPCR,and Western blotting were used to assess the osteogenic potential of the treated cells.In a rat model of bilateral ovariectomy(OVX)with femoral condylar bone defect,PHZ-1,PHZ-2,PHZ-3 or PLA/HA scaffold was implanted into the bone defect,and bone repair was observed using a microCT scanner and histological staining at 6 and 12 weeks.Results DLS,PXRD,SEM,FTIR,and EDS confirmed successful synthesis of 10-nm ZCP and efficient nZCP loading in the scaffold.PHZ-2 and PHZ-3 had significantly greater compression strength than PLA/HA.ICP-MS showed that Zn2+ release from PHZ-1,PHZ-2 and PHZ-3 were all optimal for promoting osteogenesis.In rat BMSCs,all the 4 scaffolds showed good biocompatibility,and their extracts enhanced ALP activity and extracellular matrix mineralization and promoted expressions of ALP,RUNX2,and OCN in the cells.In the rat models,nZCP in the implants improved bone graft integration at 6 weeks,and PHZ-2 and PHZ-3 more effectively induced new bone formation at 12 weeks(P<0.05).Conclusion PHZ scaffold is capable of stable Zn2+ release to promote osteoporotic bone defect healing,and PHZ-2 and PHZ-3 scaffolds with nZCP mass fraction of 4.5%-7.5%have better osteogenic activity.

Objective To investigate the release kinetics of Zn2+ from nZCP-loaded polylactic acid/hydroxyapatite(PLA/HA)composite scaffold(PHZ)and determine the optimal nZCP content in the scaffold.Methods The particle size of nZCP was measured by DLS measurement,and PXRD,FTIR,and SEM were used to characterize the scaffolds and nZCP distribution;EDS was used to analyze element composition of the scaffold.Compression strength of the scaffold was determined,and ion release profile was investigated using ICP-MS.The biocompatibility of the materials was evaluated by CCK-8 assay and dead/alive staining of rat bone marrow stem cells(BMSCs)incubated with their aqueous extracts.ALP staining,alizarin red staining,RT-qPCR,and Western blotting were used to assess the osteogenic potential of the treated cells.In a rat model of bilateral ovariectomy(OVX)with femoral condylar bone defect,PHZ-1,PHZ-2,PHZ-3 or PLA/HA scaffold was implanted into the bone defect,and bone repair was observed using a microCT scanner and histological staining at 6 and 12 weeks.Results DLS,PXRD,SEM,FTIR,and EDS confirmed successful synthesis of 10-nm ZCP and efficient nZCP loading in the scaffold.PHZ-2 and PHZ-3 had significantly greater compression strength than PLA/HA.ICP-MS showed that Zn2+ release from PHZ-1,PHZ-2 and PHZ-3 were all optimal for promoting osteogenesis.In rat BMSCs,all the 4 scaffolds showed good biocompatibility,and their extracts enhanced ALP activity and extracellular matrix mineralization and promoted expressions of ALP,RUNX2,and OCN in the cells.In the rat models,nZCP in the implants improved bone graft integration at 6 weeks,and PHZ-2 and PHZ-3 more effectively induced new bone formation at 12 weeks(P<0.05).Conclusion PHZ scaffold is capable of stable Zn2+ release to promote osteoporotic bone defect healing,and PHZ-2 and PHZ-3 scaffolds with nZCP mass fraction of 4.5%-7.5%have better osteogenic activity.

Objective:To investigate the effect of total alkaloid of harmaline (TAH) on inducing cellular autophagy and degradating of neurotoxic proteins Tau and α-synuclein (α-Syn).Methods:(1) The in vitro cultured PC12 cells were divided into blank control group, and 1, 2.5, 5, 10, 20 and 50 μg/mL TAH groups, respectively; and they were treated with 0, 1, 2.5, 5, 10, 20 and 50 μg/mL TAH for 24 h; cell morphology and number were observed, and cell survival rate was determined by MTT assay. (2) PC12 cells were divided into blank control group, rapamycin group, and 1, 2.5, 5, 10 and 20 μg/mL TAH groups; these cells were treated with same amount of solvent, 50 nmol/L autophagy activator rapamycin, and 1, 2.5, 5, 10 and 20 μg/mL TAH for 4 h, respectively, and the number of autophagosomes was detected by immunofluorescent staining. (3) PC12 cells were divided into blank control group, rapamycin group, and 10 μg/mL TAH group; these cells were treated with same amount of solvent, 50 nmol/L rapamycin, and 10 μg/mL TAH for 4 h; the protein expression levels of p62 and microtubule-associated protein 1 light chain 3 II (LC3-II) was detected by Western blotting. (4) PC12 cells were divided into blank control group, chloroquine group, TAH group, and TAH+chloroquine group; these PC12 cells were treated with 50 nmol/L autophagy inhibitor chloroquine, 10 μg/mL TAH, and 10 μg/mL TAH+50 nmol/L chloroquine for 4 h, respectively; the LC3-II protein expression was detected by Western blotting. (5) PC12 cells were divided into TAH group and blank control group; 10 μg/mL TAH and same amount of solvent were given to each group for 4 h, and then, the phosphorylated mammalian target of rapamycin (p-mTOR) and phosphorylated 70-KD ribosomal protein S6 kinase (p-P70S6K) protein expression levels were detected by Western blotting. (6) Tet on HEK293 cells with Tau-green fluorescent protein (GFP) overexpression were divided into blank control group, TAH group, doxycycline group, doxycycline+TAH group, doxycycline+TAH+3-MA group, and doxycycline+TAH+chloroquine group. Cells in the later 4 groups were treated with 200 ng/mL Tet system inducer doxycycline for 24 h; cells in the blank control group were treated with same amount of solvent, those in the TAH group were treated with 10 μg/mL TAH, and cells in the latter 3 groups were treated with 10 μg/mL TAH, 10 μg/mL TAH+5 mmol/L 3-MA, and 10 μg/mL TAH+50 nmol/L chloroquine, respectively, for 24 h; the changes of green fluorescence intensity of these cells were observed under laser confocal microscope. The Tau-GFP and LC3-II protein expression levels were detected by Western blotting. (7) HEK293 cells with stable α-Syn expression were divided into blank control group, chloroquine group, TAH group and TAH+chloroquine group; these cells were treated with same amount of solvent, 50 nmol/L chloroquine, 10 μg/mL TAH and 10 μg/mL TAH+50 nmol/L chloroquine for 24 h, respectively; the α-Syn and LC3-II protein expression levels were detected by Western blotting. Results:(1) As compared with that in the blank control group, the cell survival rate in 20 and 50 μg/mL TAH groups was significantly lower, and that in the 50 μg/mL TAH group was statistically lower than that in 20 μg/mL TAH group ( P<0.05). (2) As compared with that in the blank control group, the number of autophagosomes in rapamycin group, and 10 and 20 μg/mL TAH groups was significantly increased, and that in 10 μg/mL TAH group was statistically higher than that in 20 μg/mL TAH group ( P<0.05); 10 μg/mL TAH group was selected for subsequent experiments. (3) As compared with the blank control group, the rapamycin group and TAH group had significantly decreased P62 protein expression and significantly increased LC3-II protein expression ( P<0.05). (4) As compared with that in the blank control group, the LC3-II protein expression in the chloroquine group, TAH group and TAH+chloroquine group was significantly increased, and LC3-II protein expression in TAH+chloroquine group was statistically higher than that in chloroquine group ( P<0.05). (5) The p-mTOR and p-p70S6K expression levels in the TAH group were significantly decreased as compared with those in the blank control group ( P<0.05). (6) The Tau-GFP protein expression in doxycycline group was significantly increased as compared with that in the blank control group ( P<0.05); that in doxycycline+TAH group was significantly decreased as compared with that in the doxycycline group ( P<0.05); that in the doxycycline+TAH+3-MA group and doxycycline+TAH+chloroquine group was statistically increased as compared with that in doxycycline+TAH group ( P<0.05). The LC3-II protein expression in the TAH group was significantly increased as compared with that in the control group, that in the doxycycline+TAH group was significantly increased as compared with that in the doxycycline group, that in the doxycycline+TAH+3-MA group was significantly decreased as compared with that in the doxycycline+TAH group, and that in doxycycline+TAH+ chloroquine group was significantly increased as compared with that in the doxycycline+TAH group ( P<0.05). Conclusion:TAH may activate autophagy by inhibiting the mTOR/p70S6K signaling pathway, which in turn promotes the degradation of neurotoxic proteins Tau and α-Syn.

Objective:To evaluate the effect of sufentanil on activation of Schwann cells after peripheral nerve injury in mice.Methods:Eighty healthy pathogen-free male Balb/c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=20 each) using a random number table method: peripheral nerve injury group (group PNI), high dose sufentanil group (group H), medium dose sufentanil group (group M) and low dose sufentanil group (group L). The model of unilateral sciatic nerve transaction was established in ketamine-anesthetized mice.Immediately after establishment of the model, sufentanil 10, 5 and 2.5 μg/kg was injected intraperitoneally once a day for 3 consecutive days in H, M and L groups, respectively, while the equal volume of normal saline was given instead in group PNI.Sciatic function index (SFI) was calculated at 4, 8 and 12 weeks after establishment of the model.At 2, 4, 8 and 12 weeks, 5 mice in each group were sacrificed, and segments of the injuried ipsilateral sciatic nerve were removed for examination of the ultrastructure of the sciatic nerve (with a transmission electron microscope) and for detection of the expression of glial fibrillary acidic protein (GFAP) of sciatic nerve (by immunohistochemistry). Results:Compared with group PNI, SFI was significantly increased, and the expression of GFAP was up-regluated at each time point after establishment of the model in H and M groups ( P<0.05) and no significant change was found in SFI and GFAP expression after establishment of the model in group L ( P>0.05). Compared with group L, SFI was significantly increased, and GFAP expression was up-regluated in H and M groups ( P<0.05). There was no significant difference in SFI and GFAP expression between group H and group M ( P>0.05). The thickness of myelin lamellae was dense, and the proliferation of Schwann cells was not marked in H and M groups.The thickness of myelin lamellae was thin, and the proliferation of Schwann cells was marked in L and MO groups. Conclusion:The mechanism by which sufentanil improves repair after peripheral nerve injury may be related to promoting activation of Schwann cells in mice.