ObjectiveTo determine the subcellular localization of exogenous human papillomavirus type 16 E6 protein(HPV16 E6) and hDaxx in HeLa cells and their effects on tumor necrosis factor (TNF)-α-induced apoptosis.MethodsHeLa cells were transfected with plasmids pDsRed-monomer-C1/HPV16 E6,pEGFP-CI/hDaxx,pEGFP-C1 and pDsRed-monomer-C1 respectively.Subsequently,Western blot was carried out to quantify the expression of fusion proteins DsRed-HPV16E6 and EGFP-hDaxx in transfected cells,and laser scanning confocal microscopy to observe the subcellular distribution of HPV16 E6 protein and hDaxx.Some HeLa cells were divided into 5 groups:untransfected (control group),untransfected and treated with TNF-α(TNF-ot group),transfected with pcDNA3.1 (-) and treated with TNF-α(empty vector group),transfected with pcDNA3.1 (-)/HPV16 E6 and treated with TNF-α (HPV16 E6 group),cotransfected with pcDNA3.1(-)/HPV16 E6 and pcDNA3.1 (-)/hDaxx and treated with TNF-α (cotransfected group).After additional culture,the cells were collected and subjected to flow cytometry(FCM) to evaluate the apoptosis of cells as well as spectrophotometry to determine the relative activity of Caspase-8 and Caspase-3.ResultsWestern blot showed that both DsRed-HPV16 E6 and EGFP-hDaxx were expressed in HeLa cells.In Hela cells transfected with pDsRedmonomer-C1/HPV16 E6 or pEGFP-C1/hDaxx alone,the red fluorescence of HPV16 E6 was observed in the nucleus and cytoplasm,while the green fluorescence of hDaxx only in the nucleus; in those cotransfected with pDsRed-monomer-C1/HPVl6 E6,HPV16 E6 and hDaxx proteins were regionally aggregated near the nuclear membrane in nuclei,and hDaxx was partly translocated from the nucleus to the cytoplasm.The apoptosis rate and relative activity of Caspase-8 and Caspase-3 were statistically lower in HPV16 E6 group than in the empty vector group and cotransfected group(21.4％ ± 1.1％ vs.27.0％ ± 0.9％ and 32.5％ ± 2.1％,0.057 ± 0.003 vs.0.092 ±0.012 and 0.109 ± 0.013,0.054 ± 0.006 vs.0.093 ± 0.005 and 0.110 ± 0.004,all p＜ 0.01).Conclusions HPV16 E6 protein induces the partial translocation of hDaxx from the nucleus into the cytoplasm and colocalizes with hDaxx in the cells.The apoptosis of HeLa cells induced by TNF-α can be suppressed by HPV16 E6 protein,while the overexpression of hDaxx can attenuate the suppressing effect of HPV16 E6 protein on apoptosis in Hela cells.

Objective To study the effects of Xiaoer-Chaigui-Tuire granules in the treatment of mycoplasma pneumoniae pneumonia (MPP). Methods A total of 200 children with mycoplasma pneumoniae pneumonia which conformed to the standard of admission were randomly divided into the control group (n=100) and the treatment group (n=100). The control group were treated with azithromycin, and the treatment group were treated with Xiaoer-Chaigui-Tuire granules combined with azithromycin. The temperature recovery time, cough disappeared time, asthma disappeared time, lungs rale disappeared time of the two groups were observed and compared. The serum IgG, IgM and IgA levels were detected by ELISA and the CD3, CD4, CD8 cell ratio of peripheral blood were detected by flow cytometry. Results The total effect rate of the treatment group was 95.0％ (95/100), significantly higher than 80.0％ (80/100) of the control group (χ2=10.286, P=0.001). The fever disappeared time (2.5 ± 0.3 d vs. 4.2 ± 0.9 d, t=17.004), cough disappeared time (3.0 ± 0.6 d vs. 4.5 ± 0.5 d, t=19.861), asthma disappeared time (3.1 ± 1.1 d vs. 4.0 ± 1.3 d, t=5.046), lungs rale disappeared time (4.2 ± 1.0 d vs. 8.4 ± 1.5 d, t=22.988) and of the treatment group were significantly lower than those of the control group (P<0.01). After treatment, the IgG (9.47 ± 1.63 g/L vs. 7.05 ± 1.46 g/L, t=11.059), IgM (1.87 ± 0.52 g/L vs. 1.53 ± 0.48 g/L, t=4.804) and IgA (1.79 ± 0.61 g/L vs. 1.20 ± 0.53 g/L, t=7.301) of the treatment group were significantly higher than those of the control group (P<0.01). After treatment, the blood CD3 levels (78.83％ ± 5.20 ％ vs. 57.48％ ± 4.09％, t=32.271), CD4 levels (40.27％ ± 3.42％ vs. 30.54％ ± 3.27％, t=20.563) of the treatment group were significantly higher than the control group, and the blood CD8 levels (21.15％ ± 2.90％ vs. 30.14％ ± 3.85％, t=18.651) of the treatment group were significantly lower than the control group (P<0.01). There was no significant difference in the incidence of adverse events between two groups (χ2=0.649, P=0.421). Conclusions The Xiaoer-Chaigui-Tuire granules combined with azithromycin for children with MPP has a good efficacy and low adverse events. They could relieve the clinical symptoms and improve the children's immune function.

Objective:To explore the influencing factors and mechanism of job satisfaction of medical staff based on grounded theory.Methods:A total of 33 literatures published in CNKI from January 1, 2015 to December 31, 2019 were retrieved with the theme of " job satisfaction of medical staff" . By using NVivo 11.0 software, the 33 literatures were qualitatively analyzed, and the main categories influencing the job satisfaction of medical staff were obtained, and the action mechanism model was integrated on this basis.Results:The influencing factors of medical staff′s job satisfaction were summarized into 6 first-class nodes: work ecology, hospital management, basic health care, occupational cognition, individual characteristics and external environment. Among them, work ecology was the direct influence factor; hospital management was the key influencing factor, which played an important role in the work ecology; basic health care was the safeguard factor, which was affected by the work ecology; occupation cognition was the internal driving factor, individual characteristic was the internal uncontrollable factor, external environment was the external uncontrollable factor.Conclusions:In order to improve the job satisfaction of medical staff, we should optimize the work ecology, promote the innovation of hospital management, ensure the health and safety, and pay attention to the internalization of professional values.

Objective:To explore the role of continuous renal replacement therapy (CRRT) combined with extracorporeal carbon dioxide removal (ECCO 2R) in the treatment of children with respiratory failure. Methods:The clinical data of 12 children with respiratory failure who were treated with CRRT+ECCO 2R in PICU of Jinan Children's Hospital from July 2020 to August 2022 were collected and analyzed retrospectively. The outcomes and the external pipeline usage of the patients were observed, and the blood gas analysis and ventilator parameters before 1 h and after 1, 6, 12 and 24 h of the treatment were compared by one-way ANOVA with LSD post hoc correction. Results:Six patients successfully withdrew from CRRT+ECCO 2R and mechanical ventilation, three patients were transferred to ECMO treatment. Three cases died after voluntary withdrawal of treatment, and two cases died due to treatment failure. The mortality rate was 41.7%. After continuous treatment of CRRT+ECCO 2R for 15 to 112 h, two cases experienced extracorporeal circuit obstruction. After 1 h of treatment, PaCO 2 decreased from (64.67±24.4) mmHg to (49.42±15.54) mmHg, pH increased from (7.28±0.20) to (7.38±0.11), FiO 2 decreased from (0.85±0.13) to (0.78±0.15), PC decreased from (19.42±4.34) cmH 2O to (17.75±4.00) cmH 2O. After 24 h of treatment, PaCO 2 decreased to (39.2±5.55) mmHg, pH increased to (7.41±0.04), FiO 2 decreased to (0.46±0.11), and PC decreased to (13.8±3.36) cmH 2O, and the differences were statistically significant compared with before treatment ( P < 0.05). Conclusions:The combination of CRRT and ECCO 2R therapy can safely substitute for partial lung ventilation/perfusion function, and play a role in protecting right heart function and improving lung-kidney interaction. It can be considered as an option for extracorporeal respiratory, circulatory, and renal support, and consequently has broad prospects.

Antrodia cinnamomea is extensively used as a traditional medicine to prevention and treatment of liver cancer. However, its comprehensive chemical fingerprint is uncertain, and the mechanisms, especially the potential therapeutic target for anti-hepatocellular carcinoma (HCC) are still unclear. Using UPLC‒Q-TOF/MS, 139 chemical components were identified in A. cinnamomea dropping pills (ACDPs). Based on these chemical components, network pharmacology demonstrated that the targets of active components were significantly enriched in the pathways in cancer, which were closely related with cell proliferation regulation. Next, HCC data was downloaded from Gene Expression Omnibus database (GEO). The Cancer Genome Atlas (TCGA) and DisGeNET were analyzed by bioinformatics, and 79 biomarkers were obtained. Furtherly, nine targets of ACDP active components were revealed, and they were significantly enriched in PI3K/AKT and cell cycle signaling pathways. The affinity between these targets and their corresponding active ingredients was predicted by molecular docking. Finally, in vivo and in vitro experiments showed that ACDPs could reduce the activity of PI3K/AKT signaling pathway and downregulate the expression of cell cycle-related proteins, contributing to the decreased growth of liver cancer. Altogether, PI3K/AKT-cell cycle appears as the significant central node in anti-liver cancer of A. Cinnamomea.