Objective To construct a recombinant eukaryotic expression vector of human tumor suppressor p53-binding protein 2(TP53BP2)and transfect human embryonic kidney Expi293F cells.High-purity recombinant human full-length TP53BP2 protein was obtained and its biological activity was identified.Methods The TP53BP2 gene sequence was queried on the UniProt website,and the Expi293F expression system was optimized.The TP53BP2 gene was connected to pcDNA3.1(+)-P2A-eGFP vector by homologous recombination,and identified by double enzyme digestion and sequencing.Transect pcDNA3.1(+)-P2A-eGFP-TP53BP2 plasmid into Expi293F cells of Polyethylenimine(PEI),observe the transfection efficiency with a fluorescence microscope,collected cells from the experiment group and control group.The expression level of TP53BP2 recombinant protein was detected by Western blot(WB).Protein was purified by His label purification kit and Superdex 20010/300 GL chromatographic column.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis.The purified recombinant protein was identified by SDS-PAGE.Combining recombinant human full-length TP53BP2 protein with p65 protein was investigated for Co-immunoprecipitation(Co-IP)precipitation.Recombinant human full-length TP53BP2 protein was co-localized with p65 protein by Immunofluorescence(IF).The surface plasmon resonance(SPR)technique was used to detect the interaction between purified recombinant human full-length TP53BP2 protein and TP53BP2 antibody.Results The recombinant plasmid pcDNA3.1(+)-P2A-eGFP-TP53BP2 was successfully constructed by sequencing and double digestion.The fluorescence microscopy results showed that the transfection efficiency was about 60%.WB showed that the TP53BP2 protein was overexpressed in Expi293F cells,which proved that transfection was successful.SDS-PAGE results showed that the purity of the purified recombinant protein was above 90%,which proved that the purification was successful.Co-IP results showed that the TP53BP2 could interact with p65 protein.The results of IF showed that His tag protein,TP53BP2 protein,and p65 protein were co-located,indicating the interaction between the three proteins.SPR results showed that the purified TP53BP2 recombinant protein had good binding activity with the TP53BP2 antibody.These results all prove that the recombinant human full-length TP53BP2 protein has biological activity.Conclusion The eukaryotic expression vector of TP53BP2 gene was successfully constructed and the recombinant full-length human TP53BP2 protein with biological activity was successfully expressed in human embryonic kidney Expi293F cells.It lays a foundation for further study on the structure and function of TP53BP2.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Silicosis, one of the most serious occupational diseases in the world, is a complex pathological process with multi-cell involvement and multi-factor regulation, and its pathogenesis has not been fully elucidated.Protein phosphatase 2A (PP2A) regulates tumor signaling pathways, cell development and cell cycle.The regulatory subunit B of PP2A binds to the core enzyme, resulting in tissue expression specificity and substrate specificity of the PP2A holoenzyme complex.Protein phosphatase 2A regulatory subunit B"α (PPP2R3A) is a subunit of PP2A regulatory subunit B", which is a regulator of cell proliferation.However, the role of PPP2R3A in pulmonary fibrosis is still unclear.In this study, the pulmonary fibrosis model was established by endotracheal infusion of silica (SiO2, 250 mg/kg).Human pulmonary fibroblast-adult cells (HFP-a) were stimulated with 5 ng/mL TGF-β1 to construct fibro-related cell models.The transcription level of Ppp2r3a was detected by qRT-PCR assay.Immunofluorescence and Western blot experiments were performed to detect protein levels.Cell viability was detected by CCK-8 assay.The cell migration ability was detected by scratch test.Experimental results showed that silica nodules and collagen deposition were obvious in the SiO2 group, and the expression of PPP2R3A in lung fibroblasts increased, which could affect cell viability and migration ability, and may promote the progression of pulmonary fibrosis by regulating the expression of p53 signaling pathways.This study provides a new idea for the prevention and treatment of pulmonary fibrosis.

Background Silicosis is caused by long-term inhalation of large amounts of free silica (SiO₂) particles, and exploring its mechanism can provide new directions for the treatment of silicosis fibrosis. Objective To investigate the expression and role of fatty acid binding protein 5 (FABP5) in a silica-induced silicosis model. Methods In combination with the results of single-cell transcriptome sequencing, the expression pattern of FABP5 in mouse alveolar epithelial cells was explored by bioinformatic analysis, and the distribution pattern of fabp5 was detected by spatial transcriptomics. An in vivo model of silicosis was established by intratracheal injection with SiO₂ into mice and four groups were set up: normal saline (NS) 7 d group, NS 56 d group, SiO₂ 7 d group, and SiO₂ 56 d group. An in vitro model of silicosis was established in SiO₂-treated mouse lung epithelial cell line (MLE-12). At the whole animal level, the marker of epithelial cells (E-Cad) and the protein level of FABP5 were detected by tissue immunofluorescence assay; in vitro, the changes of fabp5 mRNA expression and protein level in MLE-12. Results The results of single-cell transcriptome sequencing and spatial transcriptome sequencing showed that the mRNA expression of fabp5 was upregulated in type II alveolar epithelial cells in the focal area of silicosis in mice, accompanied by elevated tissue immunofluorescent protein levels, and there was co-localization of E-CAD. Meanwhile, SiO₂ stimulation induced a 1.58-fold increase in fabp5 mRNA expression and a 2-fold increase in protein levels in MLE-12 cells, with significant differences (P<0.05). Conclusion The protein level of FABP5 is increased in alveolar epithelial cells in a pulmonary fibrosis model, suggesting that FABP5 may be involved in the pathological process of epithelial cells in pulmonary fibrosis.

BACKGROUND: The study aimed to explore the factors influencing protective behavior and its association with factors during the post-COVID-19 period in China based on the risk perception emotion model and the protective action decision model (PADM). METHODS: A total of 2830 valid questionnaires were collected as data for empirical analysis via network sampling in China. Structural equation modeling (SEM) was performed to explore the relationships between the latent variables. RESULTS: SEM indicated that social emotion significantly positively affected protective behavior and intention. Protective behavioral intention had significant direct effects on protective behavior, and the direct effects were also the largest. Government trust did not have a significant effect on protective behavior but did have a significant indirect effect. Moreover, it was found that government trust had the greatest direct effect on social emotion. In addition, we found that excessive risk perception level may directly reduce people's intention and frequency of engaging in protective behavior, which was not conducive to positive, protective behavior. CONCLUSION In the post-COVID-19 period, theoretical framework constructed in this study can be used to evaluate people's protective behavior. The government should strengthen its information-sharing and interaction with the public, enhance people's trust in the government, create a positive social mood, appropriately regulate people's risk perception, and, finally, maintain a positive attitude and intent of protection.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; COVID-19/prevention & control* ; China/epidemiology* ; Communicable Disease Control ; Cross-Sectional Studies ; Emotions ; Female ; Health Behavior ; Humans ; Intention ; Latent Class Analysis ; Male ; Middle Aged ; SARS-CoV-2 ; Social Behavior ; Surveys and Questionnaires ; Trust ; Young Adult

Objective To analyze the documents regarding the occupational commitment of nurses in China to provide reference for nursing managers. Methods Literature analysis was applied. Paper on occupational commitment of nurses in China from 2002 to 2016 were collected and periodicals collection, authors,scientific research funding,distribution of the research objects,sampling methods and related factors to occupational commitment were analyized. Results A total of 25 articles were collected. The research objects were mainly from general hospitals,Chinese medicine hospitals,psychiatry and community. The main research content was influencing factors analysis. No intervention research and lack of randomization were the existing problems. Conclusions Rsearch on occupational commitment of nurses has been valued in China while the research process needs to be enhanced because of lack of intervention research.

Objective To study the skills and effects of endoscopic retrograde appendicitis therapy (ERAT) in treating patients with uncomplicated acute appendicitis .Methods We enrolled 21 patients with suspected acute appendicitis who then underwent emergent ERAT between October 2014 and January 2015 .The data of treatment were collected and the operative skills and effects of ERAT were analyzed . Results ERAT was completed successfully in all the patients ,resulting in a success rate of 100% .Mean operation time of ERAT was (49 .7 ± 18 .2) min and mean hospital stay was (3 .3 ± 1 .6)d .Cannulation of the appendix lumen was the most critical step of ERAT ,and cannulation time [(5 .7 ± 4 .9)min , P< 0 .05] was shortened significantly by the use of LoopTip guidewire . Fourteen patients with intraluminal appendicoliths (7 of massive appendicoliths , 4 of sand‐like appendicoliths and 3 of sand‐like appendicoliths with luminal stenosis ) underwent endoscopic lithotomy successfully with balloon or basket ,with the success rate of 100% .One patient who presented perforation after appendicolith removal by basket was cured with conservative treatment .Appendix stent was inserted ,then pulled out after 1 week in 9 patients ,while no complaint or complication of the stent was observed .Operation time of ERAT shortened with the increase of case number .Conclusion ERAT is an effective and safe therapy for treating patients with uncomplicated acute appendicitis .The high success rate and safety of ERAT will be achieved by selecting suitable instruments for cannulation and appendicolith removal ,deciding suitable indications for stenting ,and accumulating of operative cases .

Objective To study the effect and safety of endoscopic retrograde appendicitis therapy (ERAT) in treating patients with uncomplicated acute appendicitis. Methods Patients with uncomplicated acute appendicitis were enrolled and divided into ERAT group and LA group received laparoscopic appendectomy. Then compare treat-ment condition, complications and follow-up of the two groups. Results ERAT were completed successfully in all the patients in ERAT group, while one patient underwent a reversion to open appendectomy for technical difficulties in LA group. Mean operative time was (49.7 ± 18.2) min for ERAT group and (68.9 ± 25.9) min for LA group (P <0.05). Fever relief time (1.3 ± 0.5) d, WBC normalization time (2.0 ± 0.9) d, mean bed time (0.1 ± 0.2) d and mean hospital stay (3.3 ± 1.6) d for ERAT group were significantly lower than LA group (P <0.05). 14 patients with intra-luminal appendicoliths (7 of massive appendicoliths, 4 of broken appendicoliths and 3 of broken appendicoliths with luminal stenosis) underwent endoscopic lithotomy successfully in ERAT group, resulting in a success rate of 100.00%. One patient presented perforation after ERAT was cured with conservative treatment. During the follow-up of at least 1/2 year, the rate of recurrence was 10.00% in ERAT group. 1 patient (5.00%) underwent LA at the 5th month after ERAT during the follow-up. Conclusion ERAT is an effective and safe therapy in treating patients with uncomplicated acute appendicitis with advantages of minimal invasiveness and quick recovery. Uncomplicated acute appendicitis with appendicoliths and/or luminal stenosis are the most suitable indications for ERAT.

ObjectiveAfter intratympanic gadolinium administration through the tympanic membrane,three-dimensional fluid-attenuated inversion recovery magnetic resonance imaging (3D-FLAIR MRI) was performed to evaluate endolymphatic visualization and its diagnostic value in Meniere's disease.MethodsTwenty-four hours after intratympanic gadolinium administration through the tympanic membrane,19 patients with unilateral Meniere's disease diagnosed clinically underwent 3D-FLAIR and 3D-Balance-FFE imaging at 3.0 T MR scanner.The enhanced imaging of perilymphatic space in bilateral cochlea,vestibular and (or) canal were observed.Scala tympani and scala vestibule of bilateral cochlear basal turn were scored respectively.The enhanced range of bilateral vestibule and the signal intensity ratio (SIR) between the vestibule and the brain stem were measured. Wilcoxon tests and paired t tests were used.ResultsThe gadolinium appeared in almost all parts of the perilymph in cochlea,vestibular and (or) canal,so the endolymphatic space was clearly visualized on 3D-FLAIR imaging.The score of scala vestibuli between the affected side (3 cases scored 2,9 cases scored 1,7 cascs scored 0) and the healthy side ( 15 cases scored 2,2 cases scored 1,2 cases scored 0 ) were significantly different ( U =3.090,P ＜ 0.05 ).The area of enhanced vestibular were (5.77 ± 2.33) mm2 and (8.11 ± 3.32) mm2 for the affected side and the healthy side,which were significantly different ( U =3.090,P ＜ 0.05 and t =2.638,P ＜ 0.05 ).Conclusions According to 3D-Balance-FFE MRI and the enhancement of perilymphatic space,3D-FLAIR MRI with intratympanic gadolinium injection through the tympanic membrance can be used to show the border between the perilymph and the endolymph and confirm endolymphatic hydrops,thus providing radiographic evidence for the diagnosis of Meniere's disease.

Purpose To investigate the anti-inflammatory effect of Radix Rehmanniae Praeparata(RRP)on the footpad inflammation induced by complete Freund's adjuvant(CFA)in rats.Methods CFA 100 μL were injected subcutaneously to the Wistar rats at the pad of right hindfoots.19 days later,the rats were daily siven RRP water extract(0.625,1.250,2.500 g/kg·bw)or dexamethasone(0.5mg/ks·bw)intragastrically.The changes of body weight and foot volume were measured.The indexes of organ and blood were determined at the 29th day,the foot pad was removed,and routine paraffin section was performed.Results The model rats kept foot swelling and lymphocyte infiltrating,and the platelet number decreased.The other indexes were statistically insignificant when compared to the controls.RRP did not display any anti-inflammatory effect on the swollon foots,but thoracic gland and spleen indexes were rescued,and platelet number and creatinine content were increased by RRP administration in a dose-dependent manner.The anti-inflammation of dexamethasone was conspicuous,but the side effects were also significant.Conclusion RRP may be plays an adjunctive action in herbal recipes to treat rheumatoid arthritis.