Objective : To investigate the effects of resveratrol(Res) on fibroblast-like synoviocytes(FLS) in patients with rheumatoid arthritis(RA), and to explore the possible mechanism of Res inhibiting the release of inflammatory factors from FLS. Methods : FLS from RA patients were culturedin vitroand treated with different concentrations of Res(0, 20, 40, 80, 160, 320 μmol/L). The viability of FLS cells was detected by CCK-8 assay after 12 and 24 h. The contents of inflammatory factor interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in cell supernatant were detected by ELISA. The expression levels of cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS) and stimulator of interferon gene(STING) were measured by Western blot; After lentivirus infection with FLS caused the cells to overexpress cGAS, the cells were divided into Control group(blank control), cGAS group(cGAS overexpression), Res+cGAS group(Res 160 μmol/L+cGAS overexpression) and Res group(Res 160 μmol/L). The expression level of STING protein in cells of each group was determined by Western blot, the viability of FLS cells in each group was detected by CCK-8, and the contents of inflammatory factor IL-6 and TNF-α in the supernatant of cells of each group were detected by ELISA method. Results : The results of CCK-8 experiment showed that under 40, 80, 160 μmol/L Res treatment, FLS viability decreased significantly after 24 h compared with blank control group(P<0.01). ELISA results showed that the contents of IL-6 and TNF-α in cell supernatant were also significantly decreased after treatment with Res of 40, 80 and 160 μmol/L(P<0.01). Meanwhile, Western blot results showed that Res could significantly decrease the protein expression levels of STING and cGAS in FLS cells after treatment of 40, 80 and 160 μmol/L(P<0.05,P<0.01). Compared with the Control group, the expression level of STING protein in FLS increased after overexpression of cGAS(P<0.05); compared with the Res group, the content of inflammatory factors in the supernatant of FLS and the expression level of STING protein in FLS significantly increased after overexpression of cGAS(P<0.01,P<0.05). Conclusion The appropriate concentration of Res can inhibit the release of inflammatory cytokines in FLS cells, which may be related to the blocking of cGAS-STING signaling pathway.

AIM: To compare the agreement between the ARK Biometer Combo and OA 2000 in patients wearing orthokeratology lenses.METHODS: A prospective study. A total of 148 patients(148 eyes)who were wearing orthokeratology lenses and returned for follow-up at the Shanghai Eye Disease Prevention and Treatment Center from August to September 2024 were included. Biometric measurements were performed using both the ARK Biometer Combo and OA 2000. Parameters including axial length(AL), corneal central thickness(CCT), anterior chamber depth(ACD), lens thickness(LT), corneal curvature(Kf and Ks), astigmatism(AST), white-to-white corneal diameter(WTW)and pupil diameter(PD)were obtained. Differences in measurement parameters between the two biometers were compared, and agreement was assessed.RESULTS: There were no statistically significant differences in the measurements of Kf, Ks and AST between the two biometers(P>0.05). Statistically significant differences were found in the measurements of AL, CCT, ACD, LT, WTW and PD(t=2.559, P=0.012; t=16.771, P<0.0001; t=4.749, P<0.0001; t=-15.212, P<0.0001; t=-14.915, P<0.0001; t=-2.402, P=0.018). ICC ranged from 0.615 to 0.999. Bland-Altman analysis showed that the maximum absolute values of the 95% limits of agreement(LoA)of AL, CCT, ACD, LT, Kf, Ks, AST, WTW and PD were 0.07 mm, 35.07 μm, 0.07 mm, 0.12 mm, 0.66 D, 1.14 D, 1.00 D, 0.76 mm, and 0.98 mm, respectively.CONCLUSION: In orthokeratology patients, the ARK Biometer Combo and OA 2000 showed good agreement in measuring AL, CCT, ACD, Kf and LT, and can be used interchangeably.

OBJECTIVE: To analyze the time characteristics of the Ethos online adaptive radiotherapy (OART) process in clinical practice and provide guidance for the comprehensive optimization of each stage of adaptive radiotherapy. METHODS: The study involved 61 patients with cervical, rectal, gastric, lung, esophageal, and breast cancers who underwent Ethos OART. The mean ± standard deviation of segmental time, total time, and target volume for these patients were tracked. The time characteristics for different cancer types were evaluated, and the average time for target and organ at risk (OAR) modifications was compared with the average target volume for each cancer type. RESULTS: Cervical cancer born the longest total treatment time, while breast cancer had the shortest. For all cancer types except breast cancer, the modification time for target and OAR was the most time-consuming segment. The average time for target and OAR modifications aligned with the trend of the average target volume. CONCLUSION The total treatment time for various cancers ranges from 15 to 35 minutes, indicating room for improvement.
Humans ; Radiotherapy Planning, Computer-Assisted/methods* ; Neoplasms/radiotherapy* ; Female

Poly(ADP-ribosyl)ation (PARylation) is a specific form of post-translational modification (PTM) predominantly triggered by the activation of poly-ADP-ribose polymerase 1 (PARP1). However, the role and mechanism of PARylation in the advancement of acute kidney injury (AKI) remain undetermined. Here, we demonstrated the significant upregulation of PARP1 and its associated PARylation in murine models of AKI, consistent with renal biopsy findings in patients with AKI. This elevation in PARP1 expression might be attributed to trimethylation of histone H3 lysine 4 (H3K4me3). Furthermore, a reduction in PARylation levels mitigated renal dysfunction in the AKI mouse models. Mechanistically, liquid chromatography-mass spectrometry indicated that PARylation mainly occurred in receptor for activated C kinase 1 (RACK1), thereby facilitating its subsequent phosphorylation. Moreover, the phosphorylation of RACK1 enhanced its dimerization and accelerated the ubiquitination-mediated hypoxia inducible factor-1α (HIF-1α) degradation, thereby exacerbating kidney injury. Additionally, we identified a PARP1 proteolysis-targeting chimera (PROTAC), A19, as a PARP1 degrader that demonstrated superior protective effects against renal injury compared with PJ34, a previously identified PARP1 inhibitor. Collectively, both genetic and drug-based inhibition of PARylation mitigated kidney injury, indicating that the PARylated RACK1/HIF-1α axis could be a promising therapeutic target for AKI treatment.

OBJECTIVES: To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress. METHODS: Twenty male SD rats were randomized equally into control group and heat stress group. After exposure to 32 ℃ for 2 weeks in the latter group, the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry. In the cell experiments, cultured rat thoracic aortic endothelial cells (RTAECs) were incubated at 40 ℃ for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA (si-Bmal1) or a negative sequence. In both rat thoracic aorta and RTAECs, the expressions of Bmal1, the cell cycle proteins CDK1, CDK4, CDK6, and cyclin B1, and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting. TUNEL staining was used to detect cell apoptosis in rat thoracic aorta, and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry. RESULTS: Compared with the control rats, the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1, cyclin B1 and CDK1 in the thoracic aorta (P<0.05). In cultured RTAECs, heat stress caused significant increase of Bmal1, cyclin B1 and CDK1 protein expression levels, which were obviously lowered in cells with prior si-Bmal1 transfection. Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2. CONCLUSIONS Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells, which can be partly alleviated by suppressing Bmal1 expression.
Animals ; ARNTL Transcription Factors/genetics* ; Male ; Aorta, Thoracic/metabolism* ; Rats ; Rats, Sprague-Dawley ; Endothelial Cells/metabolism* ; Apoptosis ; Cells, Cultured ; Heat-Shock Response ; Cyclin B1/metabolism* ; CDC2 Protein Kinase/metabolism* ; Cyclins/metabolism* ; RNA, Small Interfering ; bcl-2-Associated X Protein/metabolism*

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

AIM: To observe the early retinal degeneration and activation of microglia in C57BL/6N(Crb1rd8/rd8)mice.METHODS:Totally 15 male SPF C57BL/6N mice and 15 male SPF C57BL/6J mice were raised normally, and fundus photography examinations were performed by Micron-Ⅲ at the time of 0, 4, 8, 12 wk of enrollment to calculate the number and area of retinopathy. At the end of experiment, all mice were sacrificed and the right eyeballs were removed to prepare retinal tissue slices. After HE staining, the retinal tissue morphology was observed under optical microscope while the location and level of CX3CR1 expression were detected in immunohistochemical staining. The left eyeballs were removed to isolate retina, then Western-Blot was used to analyze the expression of CD86 and CD206 proteins in retina, and the concentration of IL-1β, IL-6, TNF-α, IL-4 and IL-10 in retina was detected by electrochemiluminescence.RESULTS:The result of fundus photography examinations showed that the number of retinopathy in the C57BL/6N significantly increased at 4, 8, and 12 wk, and there were differences in variations compared with the C57BL/6J at the same time point(all P<0.05). In the changes in area of retinopathy, there was a difference between two groups at 12 wk(P<0.05), but no difference in variations within groups(both P>0.05). HE staining of retinal tissue showed that the retinal structure of C57BL/6N mice was abnormal, with loose and disordered cell arrangement, and the photoreceptor layer was obviously protruding to the inner side of retina with a drusen-like protrusion. The retinal structure of C57BL/6J mice was clearer, with orderly cell arrangement and no obvious abnormality. Immunohistochemical results showed that CX3CR1 was highly expressed in ganglion cell layer, inner and outer plexiform layer, photoreceptor cell layer and lesion in the retina of C57BL/6N mice, with a mean density of 0.285±0.056 in C57BL/6N and 0.189±0.084 in C57BL/6J mice(P<0.05). The results of Western-Blot showed that the expression of CD86 and CD206 in retina of C57BL/6N increased compared with that in C57BL/6J to varying degrees, and the difference of CD86 was statistically significant(P<0.05). The results of cytokine detection showed that the level of IL-1β, TNF-α in C57BL/6N was significantly higher than that of C57BL/6J, while IL-10 was significantly lower(all P<0.05).CONCLUSION: The retinal degeneration of C57BL/6N(Crb1rd8/rd8)mice progressed slowly and gradually aggravated with age. The retinal structure of the lesion was disordered and accompanied by microglial infiltration dominated by M1 polarization.

Objective:To study the relationship between the absorptive capacity of inpatients with infectious diseases and the structure of diseases in 65 secondary and tertiary general hospitals in Beijing,and to objectively analyze the current situation of space utilization of inpatients with infectious diseases,so as to provide data support for the formulation of relevant policies.Methods:The variability of spatial absorption capacity indicators for secondary and tertiary general hospitals in 6 urban districts and 10 suburbs were compared separately,and the correlation between the spatial absorption capacity of secondary and tertiary general hospitals and the structure of disease types was visualized and analyzed using quadrant bubble charts.Results:In terms of spatial absorption capacity,there was a statistically significant difference in the proportion of patients from suburban districts treated in the secondary and tertiary general hospitals in 6 urban districts of the Beijing(P=0.003),while there was no statistically significant difference in the proportion of patients from other districts treated in the secondary and tertiary general hospitals in 10 suburbs(P=0.336).The spatial absorption capacity and disease structure of the secondary and tertiary hospitals in 6 urban districts and the tertiary hospitals in 10 suburbs showed significant correlation,while the secondary hospitals in 10 suburbs showed no significant correlation.Conclusion:The tertiary general hospitals in 6 urban districts have superior infectious disease type structure indicators,with significantly stronger spatial absorption capacity and stronger correlation between these two,which plays the function of inpatient service of difficult and severe infectious diseases.Only the district hospitals in the outer suburbs can provide inpatient services for infectious diseases,and the number of cases admitted is large,which meets the needs of inpatient diagnosis and treatment of common infectious diseases in the district.It is necessary to strengthen the investment of infectious disease medical resources and capacity building in 10 suburban districts according to the actual situation.

Objective:The service capacity of infectious diseases department in Beijing secondary and tertiary general hospitals was quantitatively analyzed to provide objective data support for the construction of infectious diseases department.Methods:The scope of infectious diseases was defined by the DRG tool,and the service capacity of medical institutions was described by the Case Mix Index(CMI)and number of DRG groups.The differences in the service capacity of infectious diseases in 67 secondary and tertiary general hospitals in Beijing from 2016 to 2020 were analyzed by data visualization and generalized linear equation,and the disease structure of different hospitals at different levels was compared by selecting representative hospitals.Results:From 2016 to 2020,the mean CMI of tertiary hospitals ranged from 0.94 to 0.97,while that of secondary hospitals ranged from 0.70 to 0.72.From 2016 to 2019,the average number of DRG groups in tertiary hospitals ranged from 26.75 to 27.79,and the average number of DRG groups in secondary hospitals ranged from 15.32 to 15.77,and the average number of DRG groups in secondary and tertiary hospitals showed a significant decline in 2020.CMI and number of DRG groups had statistical significant difference at hospital level(P＜0.001),and number of DRG groups had statistical difference in time dimension(P＜0.001).Conclusion:The infectious disease service capacity of tertiary general hospitals is obviously stronger than that of secondary general hospitals,and the infectious disease medical service capacity of large tertiary hospitals is obviously ahead,while the overall service level of secondary hospitals is low.We should give full play to the advantages of top three hospitals to build a high ground for infectious disease department construction,and combine the spatial distribution of medical resources and demand characteristics to improve the infectious disease service capacity of secondary hospitals.

Background Coal workers' pneumoconiosis (CWP) is a serious occupational lung disease and one of the prescript occupational diseases in China. Epithelial-mesenchymal transition (EMT) is involved in the diffuse fibrosis of lung tissue of pneumoconiosis patients, and its mechanism may be related to the polarization of macrophages regulated by let-7c. Objective To investigate the effect of let-7c on the regulation of macrophage polarization in EMT in rats induced by coal dust exposure with different content of free SiO₂. Methods SD rats were randomly divided into a control group, a 5% SiO₂ group, a 30% SiO₂ group, and a 99.9% SiO₂ group, with 16 rats in each group. The rats in each group were tracheally titrated with 100 μL of 20 mg·mL⁻¹ suspension (5% SiO₂, 30% SiO₂, and 99.9% SiO₂) or normal saline, respectively. Alveolar lavage fluid was collected at the ends of the 1st month and the 3rd month. The relative expression levels of M1 or M2 markers, CD86 or CD206, in alveolar macrophages (AMs) were detected by immunofluorescence. The inflammation and fibrosis of lung tissue were observed by hematoxylin-eosin (HE) staining and Masson staining. The expression levels of transforming growth factor-β1 (TGF-β1), E-cadherin, and vimentin were detected by Western blotting. The expression levels of let-7c and c/EBP-δ genes were detected by real-time fluorescence quantitative PCR. Results The HE and Masson staining results showed that compared with the control group, the degree of pulmonary fibrosis in the 5% SiO₂ group, the 30% SiO₂ group, and the 99.9% SiO₂ group gradually increased with the increase of dust exposure time. Compared with the control group, the expressions of CD86 and CD206 in the 5% SiO₂ group, the 30% SiO₂ group, and the 99.9% SiO₂ group gradually increased at the end of the 1st month (F=330.904, 146.801, P<0.05), and the expression of CD86 in each group decreased gradually at the end of the 3rd month (F=331.781, P<0.05), but the expression of CD206 increased (F=1164.190, P<0.05). At the end of the 1st month, the expressions of TGF-β1 (F=8.847, P<0.05) and vimentin (F=13.275, P<0.05) gradually increased, and the expression of E-cadherin (F=6.253, P<0.05) gradually reduced in the 5% SiO₂ group, the 30% SiO₂ group, and the 99.9% SiO₂ group. At the end of the 3rd month, the expressions of TGF-β1 (F=16.833, P<0.05) and vimentin (F=55.021, P<0.05) increased, and the expression of E-cadherin (F=12.790, P<0.05) gradually decreased in all groups. The PCR results showed that compared with the control group, the expression of let-7c mRNA in the 5% SiO₂ group, the 30% SiO₂ group, and the 99.9% SiO₂ group increased at the ends of the 1st month and the 3rd month (F=11.251, 28.136, P<0.05). The expression of c/EBP-δ mRNA decreased in all groups at the ends of the 1st month and the 3rd month (F=49.204, 177.090, P<0.05). Conclusion In response to dust stimulation, let-7c promotes EMT by modulating macrophage polarization, which is involved in the formation of pulmonary fibrosis and thus influences the progression of CWP .