Objective To investigate the effects of fractionated low-dose ionizing radiation (LDIR) on the ferroptosis in human bronchial epithelial (HBE) cells as well as the associated differentially expressed genes (DEGs), biological processes, and signaling pathways. Methods HBE cells were exposed to different single doses of X-ray irradiation (0, 25, 50, 75, and 100 mGy) for 24, 48, and 72 h, respectively. The change in cell viability was detected by MTT assay. Cells were irradiated with 0, 25, 50, and 100 mGy X-rays 5 times, with 48 h between each irradiation and a dose rate of 50 mGy/min. Cells were harvested 24 h after irradiation for the measurement of the expression of ferroptosis-related genes SLC7A11 and GPX4 at the mRNA and protein levels, cellular iron content, and the expression of FTH1 and FTL mRNAs. High-throughput sequencing was used to screen for the DEGs in each dose group, followed by Gene Ontology-Biological Process (GO-BP) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and Gene Set Enrichment Analysis (GSEA). Results Compared with the control group, single-dose LDIR significantly increased cell proliferation at 75 mGy after 24 h (P < 0.05), at 50, 75, and 100 mGy after 48 h (P < 0.05), and at 75 and 100 mGy after 72 h (P < 0.05). Compared with the control group, at the end of the fifth fractionated LDIR, SLC7A11 and GPX4 mRNAs decreased at all doses (P < 0.05), SLC7A11 protein decreased at all doses, GPX4 protein decreased at 25 and 100 mGy, iron content increased at all doses, and FTH1 and FTL mRNAs decreased at all doses (P< 0.05). Sequencing analysis identified 248, 30, and 291 DEGs and 10, 2, and 9 ferroptosis-associated genes at the three doses compared to the control. Gene Ontology-Biological Process analysis showed that DEGs were mainly enriched in biological processes such as response to lipids, cell death, and response to unfolded proteins. Kyoto Encyclopedia of Genes and Genomes analysis showed that DEGs were mainly enriched in the JAK-STAT signaling pathway, lipids and atherosclerosis, ferroptosis, protein processing in the endoplasmic reticulum, and FoxO signaling pathway. Gene set enrichment analysis showed that DEGs were mainly enriched in ferroptosis, fatty acid degradation, and glutathione metabolism. Conclusion Fractionated low-dose radiation induced ferroptosis in HBE cells, and DEGs were predominantly enriched in biological processes and signaling pathways related to inflammation, ferroptosis, and endoplasmic reticulum stress.

Objective To investigate the role of the O-6-methylguanine-DNA methyltransferase (MGMT) gene-3′ untranslated region (UTR) microRNA-associated single nucleotide polymorphism (miRSNP) (rs7896488 G>A) in affecting miR-4297-targeted modulation of MGMT in senescence of lung cells with polymorphic genotypes induced by fractionated low dose ionizing radiation (LDIR). Methods i) MiRSNPs were predicted and screened using bioinformatics, and DNA from two types of lung cells, A549 cells and human bronchial epithelioid cells (HBE cells), was extracted for target gene sequencing. After co-transfection of pGL3c-MGMT-3′UTR-rs7896488 G>A reporter gene recombinant plasmid, pRL-TK Vector with micrON mimic NC #22 or micrON hsa-miR-4297 mimic (set up as the mimic NC group and the miR-4297 mimic group) in these two types of lung cells, dual luciferase reporter gene assay was performed. The relative expression of MGMT mRNA was detected by real-time fluorescence quantitative polymerase chain reaction, and the relative expression of MGMT protein was detected by Western blotting. ii) These two types of lung cells were randomly divided into the control group and irradiation group, which received either 0 or 100 mGy X-rays irradiation seven times. After irradiation, the cells were transfected with either micrON mimic NC #22 or micrON hsa-miR-4297 mimic, resulting in mimic NC + control group, miR-4297 mimic + control group, mimic NC + irradiation group, and miR-4297 mimic + irradiation group. Cells were collected for senescence-associated-β-galactosidase (SA-β-Gal) staining, and the relative expression of matrix metalloproteinase-9 (MMP-9) and chemokine (C-X-C motif) ligand-1 (CXCL-1) proteins was detected via Western blotting. Results i) The rs7896488 G>A was the miRSNP located in the conserved binding region targeted by miR-4297 in the MGMT gene 3′UTR. A549 cells were the rs7896488 GG wild-type homozygous genotype, while HBE cells were the rs7896488 GA heterozygous mutant genotype. In the miR-4297 mimic group, A549 and HBE cells carrying the rs7896488 G allele showed significantly lower dual-luciferase activity compared with that in the mimic NC group (both P<0.01). However, there was no significant difference in dual-luciferase activity between the two groups in both A549 and HBE cells carrying the rs7896488 A allele (both P>0.05). The relative expression levels of MGMT mRNA and MGMT protein of A549 cells in the miR-4297 mimic group were lower than those in the mimic NC group (both P<0.05). However, there was no significant difference in MGMT mRNA and MGMT protein of HBE cells between these two groups (both P>0.05). ii) The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of A549 cells in the miR-4297 mimic+irradiation group were higher than those in the mimic NC + control group, the miR-4297 mimic + control group, and the mimic NC + irradiation group (all P<0.05). The relative activity of SA-β-Gal and the relative expression of MMP-9 and CXCL-1 proteins of HBE cells in the miR-4297 mimic + irradiation group were higher than those in the mimic NC + control group and the miR-4297 mimic + control group (all P<0.05), while there was no significant difference compared with those in the mimic NC + irradiation group (all P>0.05). Conclusion MGMT-3′UTR-miRSNP rs7896488 G>A plays a role in LDIR-induced senescence of lung cells with different polymorphic genotypes by affecting miR-4297-targeted regulation of MGMT.

Objective To explore the differentially expressed mRNAs and related biological processes and pathways in fractional low-dose ionizing radiation (LDIR)-induced senescence of normal human bronchial epithelial (HBE) cells by high-throughput mRNA sequencing and bioinformatics techniques. Methods Senescence-associated β-galactosidase staining and senescence-associated secretion phenotype gene mRNA and protein expression levels were measured at 24 and 48 h after irradiating HBE cells 7 times at doses of 0, 50, 100, and 200 mGy, respectively. The differentially expressed genes were screened by high-throughput sequencing for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Results The senescence-positive area of fractional low-dose irradiated HBE cells increased in a dose-dependent manner (P < 0.05). The mRNA levels and protein expression of transforming growth factor-β1(TGF-β1) and matrix metalloproteinase-9(MMP-9) genes were increased in the 100 mGy × 7 and 200 mGy × 7 groups at 24 and 48 h after the end of irradiation compared with the control group. High-throughput sequencing showed that there were 882, 475, and 1205 differentially expressed mRNAs in each dose group compared with the control group. GO analysis showed that the differentially expressed mRNAs in each dose group were mainly enriched in biological processes such as cell cycle regulation, regulation of nitrogen compound metabolic process, regulation of cell division and response to stimulus. KEGG analysis showed that the differentially expressed mRNAs were mainly enriched in the pathways of cell cycle, cell senescence, and ferroptosis. Conclusion Fractional LDIR induced senescence in HBE cells, and differentially expressed mRNA-associated biological processes and pathways in senescent cells are related to cell cycle and cell senescence.

Objective To investigate the mechanism of fractionated low-dose ionizing radiation (LDIR) in the induction of EA.hy926 cell senescence. Methods EA.hy926 cells were irradiated with X-ray at 0, 50, 100, and 200 mGy × 4, respectively, and cultured for 24, 48, and 72 h. Several indicators were measured, including the levels of cellular senescence-associated β-galactosidase (SA-β-gal) staining, mRNA levels of senescence-associated cell cycle protein-dependent kinase inhibitor genes CDKN1A and CDKN2A, reactive oxygen species (ROS), total antioxidant capacity (T-AOC), and phosphorylated H2A histone family member X (γ-H2AX). Results After 4 fractionated LDIR, compared with the control group, the treatment groups showed increased nucleus area, blurred cell edge, and increased SA-β-gal positive area (P < 0.05) at 24, 48 and 72 h. After 4 fractionated LDIR, the mRNA level of CDKN1A increased in the 100 and 200 mGy × 4 groups at 24 and 48 h (P < 0.05), and CDKN2A mRNA level increased in the 100 and 200 mGy × 4 groups at 48 and 72 h (P < 0.05). The fluorescence intensity of ROS increased in treatment groups at 24, 48, and 72 h after 4 fractionated LDIR (P < 0.05). After 4 fractionated LDIR, the T-AOC level increased in the 100 and 200 mGy × 4 groups at 24 h (P < 0.05), and T-AOC level increased in all treatment groups at 48 and 72 h (P < 0.05). After 4 fractionated LDIR, γ-H2AX fluorescence intensity increased in all treatment groups at 24 h (P < 0.05), and the fluorescence intensity increased in the 100 and 200 mGy × 4 groups at 48 and 72 h (P < 0.05). Conclusion Fractionated LDIR can induce cellular senescence in EA.hy926 cells by impacting the cellular oxidation-antioxidation and oxidative damage levels, and the effects were relatively evident at 100 and 200 mGy.

Objective:To investigate the method and effect of anatomic reduction of orbital septum fat in blepharoplasty to correct the first and second degree of upper eyelid depression.Methods:From January 2019 to January 2021, the data of patients with first and second degree upper eyelid depression admitted to Beijing Anzhen Hospital, Capital Medical University were retrospectively analyzed. Orbital septum was fully exposed and cut in blepharoplasty, the orbital fat of upper eyelid central group and the medial group were dissected and seperated to form the fat flap pedicled on the medial side. The fat flap was rotated 180° to the medial side and fixed with the orbital fat of the medial group to correct the upper eyelid depression. Complications such as infection, hematoma, fat liquefaction, nodules and orthers were observed after surgery. Questionnaire was used to investigate the satisfaction of patients, with 1-4 points indicating dissatisfaction, general satisfaction, satisfaction and very satisfaction, respectively.Results:A total of 62 patients were included, all patients were female, aged 18-44 years, wit mean 27 years. All patients were bilateral upper eyelid depression. After 6 months of follow-up, the curvature of the double eyelid line was smooth, the width and narrowness of double eyelid were natural, and the upper eyelid depression was significantly improved. Satisfaction survey showed that 60 cases were very satisfied and 2 cases were satisfied. Wound infection was occurred in 1 case, and was cured after dressing change. Scar hyperplasia was observed in 2 cases, which was obviously alleviated after external treatment with anti-scar hyperplasia drugs. No fat liquefaction, nodule and other complications occured.Conclusions:For the first and second degree of upper eyelid depression, it can be corrected by anatomic reduction of orbital septal fat in blepharoplasty. The operation is simple, with fewer complication and high satisfaction.

【Objective】 To screen the sterilizing-grade filters applicable for production of human coagulation factor Ⅷ/von Willebrand factor complex(FⅧ/VWF)and study the sterilization filtration process. 【Methods】 Four sterilizing-grade filters for FⅧ/VWF were evaluated through indicators such as filtration capacity, filtration flux, recovery rate of FⅧ activity, recovery rate of VWF activity, recovery rate of VWF antigen, recovery rate of protein and VWF molecular distribution. The sterilizing-grade filter with the best filtration performance was selected for further study. The study was designed by general full-factor design to determine the appropriate filitered protein concentration and filitered speed range through evaluating the total filtered protein amount, recovery rate of protein and filtration efficiency, and then the process operation parameters was determined. 【Results】 The filtration flux of Sartobran P, Sartopore 2 XLG, Sartopore Platinum and Sartopore 2 XLI were 1.71±0.01, 1.80±0.01, 1.34±0.01, and 1.81±0.04 L·(m²)^-1·min^-1, respectively; the recovery rates (%) of FⅧ activity were 97.09±2.82, 99.22±0.99, 96.87±1.85 and 93.76±1.21, respectively; the recovery rates (%) of VWF activity were 98.12±1.42, 99.95±1.85, 94.80±1.62 and 92.09±1.67, respectively. Between Sartopore 2 XLG and Sartobran P, the difference of filtration flux (P<0.001) was statistically significant; between Sartopore 2 XLG and Sartopore Platinum, the differences of the filtration flux (P<0.001) and VWF potency recovery rate (P<0.05) were statistically significant; between Sartopore 2 XLG and Sartopore 2 XLI, the differences of FⅧ potency recovery rate (P<0.01) and VWF potency recovery rate (P<0.01) were statistically significant. The optimal process operating space of Sartopore 2 XLG was protein concentration of 0.45-0.58 mg/mL, and filtration rate of 1.48-2.95 L·(m²)^-1·min^-1. 【Conclusion】 Sartopore 2 XLG is the most suitable filter for the production of FⅧ/VWF and the DoE test proves that it has good process operation space.

Objective:To investigate the clinical effect of filling orbital septum with autologous fat granules for correction of severe upper eyelid depression.Methods:From January 2019 to January 2021, the data of patients with severe upper eyelid depression admitted to Beijing Anzhen Hospital, Capital Medical University were retrospectively analyzed. Fat granules were obtained from the abdomen or inner thigh. The middle part of the upper eyelid depression near the lower edge of eyebrow were selected as the injection point. The needle for filling was successively passed through the skin, subcutaneous fat layer, orbicularis oculi muscle, retro-orbicularis oculi fat, and the superior orbital margin. The needle was inserted into the orbital septum closedly to the superior orbital wall. Fat granules were injected into the orbital septum until the upper eyelid depression was corrected with slight fullness. Postoperative follow-up was conducted on the improvement of upper eyelid depression and the occurrence of complications in the patients. For the patients with poor correction of upper eyelid depression, autologous fat granule injection was performed again half a year later.Results:A total of 42 female patients with severe upper eyelid depression were included, aged 18-57 years with an average age of 36.8 years. Fat injection volume was 1.8-3.0 ml in each side. After 6-12 months of follow-up, 39 patients (92.9%) were significantly improved after one operation, and the other 3 patients (7.1%) were effectively improved after two fat filling. 2-3 days after operation, 4 cases of fat nodules appeared on upper eyelid, which subsided after local repeated massage by doctors. No complications such as hematoma, ptosis and embolism occurred.Conclusion:Filling the orbital septum with autologous fat granules can effectively improve the severe upper eyelid depression to attain eyelid rejuvenation.

Objective:To investigate the method and effect of retroauricular three flaps for repair of congenital earlobe defect.Methods:From March 2019 to December 2021, the data of patients with congenital earlobe defect treated with retroauricular three-flap method in Beijing Anzhen Hospital, Capital Medical University and Plastic Surgery Hospital, Chinese Academy of Medical Sciences were retrospectively analyzed. The specific method was to design three flaps in the posterior auricular region, A, B, C respectively. The area of flap A was larger 10%-20% than the defect area, and the flap A was rotated in the direction of the anterior and inferior earlobe. The flap B was designed as a rotation flap to repair the secondary defect after flap transplantation. The flap C was designed as a sliding flap to reduce the tension of the wound in the retroauricular region. Postoperative observation included flap blood flow, complications of infection, hematoma, and so on, and scar after wound healing. A questionnaire was used to investigate the satisfaction of patients or guardians, with scores from 1 to 4 indicating dissatisfaction, general satisfaction, satisfaction, and very satisfaction respectively.Results:A total of 10 patients were included, including 3 males and 7 females. Aged from 7-18 years old, with an average of 11 years old. All cases were unilateral lobe defects. Postoperative infection occurred in 1 case and small local hematoma in 1 case. All cases were cured after dressing change without flap necrosis and other complications. After 3 to 6 months of follow-up, there was no flap necrosis, the color, shape and plumpness of the repaired earlobe were good, and the scar behind the ear was concealed. The satisfaction survey showed that 8 cases had great satisfaction and 2 cases had satisfaction.Conclusion:Partial or total defects of earlobe can be repaired by retroauricular three flaps. The reconstructed earlobe has natural shape, hidden scar and satisfactory clinical effect.

Objective:To investigate the clinical effect of filling orbital septum with autologous fat granules for correction of severe upper eyelid depression.Methods:From January 2019 to January 2021, the data of patients with severe upper eyelid depression admitted to Beijing Anzhen Hospital, Capital Medical University were retrospectively analyzed. Fat granules were obtained from the abdomen or inner thigh. The middle part of the upper eyelid depression near the lower edge of eyebrow were selected as the injection point. The needle for filling was successively passed through the skin, subcutaneous fat layer, orbicularis oculi muscle, retro-orbicularis oculi fat, and the superior orbital margin. The needle was inserted into the orbital septum closedly to the superior orbital wall. Fat granules were injected into the orbital septum until the upper eyelid depression was corrected with slight fullness. Postoperative follow-up was conducted on the improvement of upper eyelid depression and the occurrence of complications in the patients. For the patients with poor correction of upper eyelid depression, autologous fat granule injection was performed again half a year later.Results:A total of 42 female patients with severe upper eyelid depression were included, aged 18-57 years with an average age of 36.8 years. Fat injection volume was 1.8-3.0 ml in each side. After 6-12 months of follow-up, 39 patients (92.9%) were significantly improved after one operation, and the other 3 patients (7.1%) were effectively improved after two fat filling. 2-3 days after operation, 4 cases of fat nodules appeared on upper eyelid, which subsided after local repeated massage by doctors. No complications such as hematoma, ptosis and embolism occurred.Conclusion:Filling the orbital septum with autologous fat granules can effectively improve the severe upper eyelid depression to attain eyelid rejuvenation.

Objective:To investigate the method and effect of retroauricular three flaps for repair of congenital earlobe defect.Methods:From March 2019 to December 2021, the data of patients with congenital earlobe defect treated with retroauricular three-flap method in Beijing Anzhen Hospital, Capital Medical University and Plastic Surgery Hospital, Chinese Academy of Medical Sciences were retrospectively analyzed. The specific method was to design three flaps in the posterior auricular region, A, B, C respectively. The area of flap A was larger 10%-20% than the defect area, and the flap A was rotated in the direction of the anterior and inferior earlobe. The flap B was designed as a rotation flap to repair the secondary defect after flap transplantation. The flap C was designed as a sliding flap to reduce the tension of the wound in the retroauricular region. Postoperative observation included flap blood flow, complications of infection, hematoma, and so on, and scar after wound healing. A questionnaire was used to investigate the satisfaction of patients or guardians, with scores from 1 to 4 indicating dissatisfaction, general satisfaction, satisfaction, and very satisfaction respectively.Results:A total of 10 patients were included, including 3 males and 7 females. Aged from 7-18 years old, with an average of 11 years old. All cases were unilateral lobe defects. Postoperative infection occurred in 1 case and small local hematoma in 1 case. All cases were cured after dressing change without flap necrosis and other complications. After 3 to 6 months of follow-up, there was no flap necrosis, the color, shape and plumpness of the repaired earlobe were good, and the scar behind the ear was concealed. The satisfaction survey showed that 8 cases had great satisfaction and 2 cases had satisfaction.Conclusion:Partial or total defects of earlobe can be repaired by retroauricular three flaps. The reconstructed earlobe has natural shape, hidden scar and satisfactory clinical effect.