The rapid emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants that evade immunity elicited by vaccination has posed a global challenge to the control of the coronavirus disease 2019 (COVID-19) pandemic. Therefore, developing countermeasures that broadly protect against SARS-CoV-2 and related sarbecoviruses is essential. Herein, we have developed a lipid nanoparticle (LNP)-encapsulated mRNA (mRNA-LNP) encoding the full-length Spike (S) glycoprotein of SARS-CoV-2 (termed RG001), which confers complete protection in a non-human primate model. Intramuscular immunization of two doses of RG001 in Rhesus monkey elicited robust neutralizing antibodies and cellular response against SARS-CoV-2 variants, resulting in significantly protected SARS-CoV-2-infected animals from acute lung lesions and complete inhibition of viral replication in all animals immunized with low or high doses of RG001. More importantly, the third dose of RG001 vaccination elicited effective neutralizing antibodies against current epidemic XBB and JN.1 strains and similar cellular response against SARS-CoV-2 Omicron variants (BA.1, XBB.1.16, and JN.1) were observed in immunized mice. All these results together strongly support the great potential of RG001 in preventing the infection of SARS-CoV-2 variants of concern (VOCs).

Objective:To explore the cleaning quality of different cleaning methods on robotically luminal apparatuses.Methods:This research innovated and invented specially cleaning holder that could support boiling after reducing pressure,and protective device for preprocessing so as to better clean robotically luminal apparatuses.A total of 600 robotic luminal apparatuses,which were used in urology surgery at The First Affiliated Hospital with Nanjing Medical University in November 2023,were selected as the research object,and they were divided into control group,experimental group 1 and experimental group 2 according to the principle of randomization and balance,with 200 apparatuses in each group.The luminal apparatuses of control group were cleaned as the cleaning process of the manufacturer's instructions,and these of experimental group 1 were cleaned through ultrasonic cleaning machine with perfusion in the cleaning process of the manufacturer's instructions,and boiling after reducing pressure was selected to clean those of experimental group 2 through specially cleaning holder that was innovated and invented,which could support boiling after reducing pressure.Both two experiment groups adopted protective device that was invented by our research patent.The visual estimation and magnifying glass(5-10 times)with light source,adenosine triphosphate(ATP)fluorescence detector,and detection device for protein residue were adopted to test and judge cleaning quality,cleaning time,and wear rate of transportation in luminal apparatus after cleaning.Results:The qualified rates of visual estimation and magnifying glass with light source in control group,experiment group 1 and experiment group 2 were respectively 94.5%,99.0%and 99.5%,and the differences of pairwise comparison among three groups were significant(x2=6.44,8.591,P<0.05).The qualified rates of the fluorescence detection and the detection for protein residue in control group were respectively 93.5%and 90%,and those in experiment group 1 were respectively 98.5%and 97%,and the differences among two groups were significant(x2=6.51,11.41,P<0.05).For the cleaning time of single apparatus,the experiment group 1 was shortest,and the experiment group 2 was longest,and there were significant differences in pairwise comparison among three groups(t=2.981,48.178,34.419,P<0.05).The cleaning time of 8 cleaned apparatuses of experiment group 1 still can keep advantage,while there was not significant difference in that between it and control group(P>0.05).The cleaning time of 12 cleaned apparatuses of experimental group 2 was shortest,and that of experiment group 1 was longest,and the differences of that among three groups were significant(t=2.743,5.292,3.177,P<0.05).The average wear rates of transportation of control group,experiment group 1 and experiment group 2 were respectively(9.5±1.8)%,(6.2±1.5)%and(5.8±1.3)%,and the average wear rates of transportation of experiment group 1 and 2 were significantly lower than that of control group,and the differences of that in pairwise comparison among three groups were significant(t=21.7,24.3,P<0.05),while the difference of that between two experiment groups was not significant(P>0.05).Conclusion:The cleaning technique of boiling after reducing pressure can play better cleaning effect for the slender lumen of robotic apparatuses,while the wear rates of experiment group 1 and 2,which use protective device for preprocessing,are better than that of control group.The full load amount of the application of the special cleaning holder of the robotic luminal apparatuses of experiment group 2 can increase by 1 time,which is high efficient and convenient.

OBJECTIVE: To elucidate the role and mechanism of microRNA-145-5p (miR-145-5p) in hypoxia-induced pyroptosis of human alveolar epithelial cells. METHODS: In vitro, human alveolar epithelial cell line BEAS-2B was cultured. Cells in the logarithmic growth phase were cultured to 80% confluence and then used for the experiment. (1) BEAS-2B cells were cultured under 1% O₂ hypoxic condition, with a normoxic control group. Western blotting was employed to detect the expressions of pyroptosis marker proteins [NOD-like receptor protein 3 (NLRP3), Gasdermin D N-terminal domain (GSDMD-N), and caspase-1] in cells cultured for 24 hours. Real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of miR-145-5p in cells cultured for 6 hours and 12 hours. (2) Cells were transfected with 30 nmol/L miR-145-5p mimic to overexpress miR-145-5p expression under normoxic condition or 30 nmol/L miR-145-5p inhibitor to suppress miR-145-5p expression under hypoxic condition. Control group and negative control group were respectively set up. After 24 hours of cell culture, Western blotting was used to detect the expressions of pyroptosis marker proteins and nuclear factor-E2-related factor 2 (Nrf2) in cells. Flow cytometry was applied to detect the level of reactive oxygen species (ROS) in cells. The target genes of miR-145-5p were predicted by miR target gene prediction software miRWalk and verified by Western blotting. (3) Under hypoxic condition, cells were transfected with 6.94 ng/μL silent information regulator 5 (Sirt5) overexpression plasmid or pretreated with 12.5 mmol/L N-acetyl-L-cysteine (NAC) as an ROS inhibitor. The empty plasmid group and control group were set up. After 24 hours of cell culture, Western blotting was used to detect the expressions of Sirt5, Nrf2, and pyroptosis marker proteins in cells. Flow cytometry was used to detect the level of ROS in cells. RESULTS: (1) Compared with the normoxic control group, the expression levels of pyroptosis marker proteins in the 24-hour hypoxia group was significantly increased, indicating that hypoxia could induce pyroptosis in BEAS-2B cells. The expression level of miR-145-5p in cells gradually increased with the extension of hypoxia induction time, indicating that hypoxia could cause the increase of miR-145-5p expression level. (2) The expression levels of pyroptosis marker proteins in cells of miR-145-5p mimic group significantly increased under normoxic condition as compared with the control and negative control groups [NLRP3 protein (NLRP3/β-actin): 1.58±0.07 vs. 1.00±0.01, 0.98±0.07, GSDMD-N protein (GSDMD-N/β-actin): 1.71±0.03 vs. 1.01±0.01, 0.85±0.03, caspase-1 protein (caspase-1/β-actin): 2.33±0.04 vs. 1.01±0.01, 1.05±0.04, all P < 0.05], Nrf2 protein expression level was significantly decreased (Nrf2/β-actin: 0.79±0.03 vs. 1.00±0.01, 1.03±0.04, both P < 0.05), ROS level was significantly up-regulated (fluorescence intensity: 1.74±0.03 vs. 1.00±0.01, 0.92±0.03, both P < 0.05). Under hypoxia condition, compared with control group and negative control group, the expression levels of pyroptosis marker proteins in miR-145-5p inhibitor group were significantly decreased [NLRP3 protein (NLRP3/β-actin): 0.21±0.04 vs. 1.70±0.02, 1.63±0.04; GSDMD-N protein (GSDMD-N/β-actin): 1.32±0.02 vs. 2.51±0.02, 2.72±0.03; caspase-1 protein (caspase-1/β-actin): 0.56±0.01 vs. 2.77±0.02, 3.12±0.03; all P < 0.05], Nrf2 protein expression level was significantly increased (Nrf2/β-actin: 1.57±0.04 vs. 1.22±0.01, 1.28±0.04, both P < 0.05), ROS level was significantly down-regulated (fluorescence intensity: 0.64±0.05 vs. 1.87±0.04, 1.70±0.07, both P < 0.05). The results indicated that miR-145-5p could promote cell pyrodeath. The predictive result of miRWalk showed that the 3' untranslated region (3'UTR) of Sirt5 had complementary base binding sites with miR-145-5p. The expression level of Sirt5 protein in cells of miR-145-5p mimic group was significantly lower than that of control group and negative control group under normoxic condition (Sirt5/β-actin: 0.59±0.03 vs. 1.00±0.01, 1.01±0.03, both P < 0.05), which verified that Sirt5 was the target gene of miR-145-5p. (3) The occurrence of pyrodeath could be partially reversed by transfection with Sirt5 overexpression plasmid or adding ROS inhibitor NAC into cells, and Sirt5 overexpression could also up-regulate Nrf2 expression and eliminate intracellular ROS. CONCLUSION In human alveolar epithelial cells, miR-145-5p can down-regulate Nrf2 by targeting Sirt5, thereby increasing ROS expression and inducing pyrodeath.
Humans ; MicroRNAs ; Pyroptosis ; Cell Hypoxia ; Alveolar Epithelial Cells/cytology* ; Cell Line ; NLR Family, Pyrin Domain-Containing 3 Protein ; Caspase 1/metabolism* ; Epithelial Cells/metabolism* ; Gasdermins ; Phosphate-Binding Proteins

Objective:To explore the cleaning quality of different cleaning methods on robotically luminal apparatuses.Methods:This research innovated and invented specially cleaning holder that could support boiling after reducing pressure,and protective device for preprocessing so as to better clean robotically luminal apparatuses.A total of 600 robotic luminal apparatuses,which were used in urology surgery at The First Affiliated Hospital with Nanjing Medical University in November 2023,were selected as the research object,and they were divided into control group,experimental group 1 and experimental group 2 according to the principle of randomization and balance,with 200 apparatuses in each group.The luminal apparatuses of control group were cleaned as the cleaning process of the manufacturer's instructions,and these of experimental group 1 were cleaned through ultrasonic cleaning machine with perfusion in the cleaning process of the manufacturer's instructions,and boiling after reducing pressure was selected to clean those of experimental group 2 through specially cleaning holder that was innovated and invented,which could support boiling after reducing pressure.Both two experiment groups adopted protective device that was invented by our research patent.The visual estimation and magnifying glass(5-10 times)with light source,adenosine triphosphate(ATP)fluorescence detector,and detection device for protein residue were adopted to test and judge cleaning quality,cleaning time,and wear rate of transportation in luminal apparatus after cleaning.Results:The qualified rates of visual estimation and magnifying glass with light source in control group,experiment group 1 and experiment group 2 were respectively 94.5%,99.0%and 99.5%,and the differences of pairwise comparison among three groups were significant(x2=6.44,8.591,P<0.05).The qualified rates of the fluorescence detection and the detection for protein residue in control group were respectively 93.5%and 90%,and those in experiment group 1 were respectively 98.5%and 97%,and the differences among two groups were significant(x2=6.51,11.41,P<0.05).For the cleaning time of single apparatus,the experiment group 1 was shortest,and the experiment group 2 was longest,and there were significant differences in pairwise comparison among three groups(t=2.981,48.178,34.419,P<0.05).The cleaning time of 8 cleaned apparatuses of experiment group 1 still can keep advantage,while there was not significant difference in that between it and control group(P>0.05).The cleaning time of 12 cleaned apparatuses of experimental group 2 was shortest,and that of experiment group 1 was longest,and the differences of that among three groups were significant(t=2.743,5.292,3.177,P<0.05).The average wear rates of transportation of control group,experiment group 1 and experiment group 2 were respectively(9.5±1.8)%,(6.2±1.5)%and(5.8±1.3)%,and the average wear rates of transportation of experiment group 1 and 2 were significantly lower than that of control group,and the differences of that in pairwise comparison among three groups were significant(t=21.7,24.3,P<0.05),while the difference of that between two experiment groups was not significant(P>0.05).Conclusion:The cleaning technique of boiling after reducing pressure can play better cleaning effect for the slender lumen of robotic apparatuses,while the wear rates of experiment group 1 and 2,which use protective device for preprocessing,are better than that of control group.The full load amount of the application of the special cleaning holder of the robotic luminal apparatuses of experiment group 2 can increase by 1 time,which is high efficient and convenient.

Objective:To understand the effect of presenteeism behavior of nurses on patient satisfaction and to explore the correlation between them.Methods:This study was a cross-sectional study. In December 2019, the convenient sampling method was adopted to select 500 in-service responsible nurses and 500 inpatients under the charge of the responsible nurses in 5 Class Ⅲ Grade A hospitals in Henan Province as the research objects. The responsible nurses were surveyed by general information questionnaire and Presenteeism Behavior Scale and patients in charge were surveyed by Inpatient Satisfaction and Experience Monitoring Scale.Results:A total of 435 valid questionnaires were collected from responsible nurses and patients in charge. The score of Presenteeism Behavior Scale of 435 nurses was (2.62±0.93) and the score of Inpatient Satisfaction and Experience Monitoring Scale of 435 patients was (4.50±0.49) , showing negative correlation ( r=-0.19, P<0.01) . Conclusions:The level of nurses' presenteeism behavior is at a moderate level and patient satisfaction is at an upper-middle level. Nurses' presenteeism behavior affects patient satisfaction. The higher the incidence of nurses' presenteeism behavior, the lower the patient satisfaction. It is recommended to reduce nurses' presenteeism behavior and improve patient satisfaction.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has become one major threat to human population health. The RNA-dependent RNA polymerase (RdRp) presents an ideal target of antivirals, whereas nucleoside analogs inhibitor is hindered by the proofreading activity of coronavirus. Herein, we report that corilagin (RAI-S-37) as a non-nucleoside inhibitor of SARS-CoV-2 RdRp, binds directly to RdRp, effectively inhibits the polymerase activity in both cell-free and cell-based assays, fully resists the proofreading activity and potently inhibits SARS-CoV-2 infection with a low 50% effective concentration (EC

Human APOBEC3G (hA3G) is a cytidine deaminase which inhibits HIV-1 replication. The HIV-1 accessory protein viral infectivity factor (Vif) counteracts with hA3G by targeting it for proteasomal degradation. In this work, we constructed and optimized molecular models of the hA3G dimer and the hA3G-Vif complex. The molecular modeling study revealed that the loop7 motif of hA3G appears on the interfaces of both the hA3G-Vif complex and the hA3G dimer. Biochemical analysis provided evidence suggesting that binding of Vif to hA3G results in steric blocking of hA3G dimerization, implying that monomeric hA3G serves as a substrate for Vif-mediated degradation. Furthermore, we presented evidence for the important roles of the loop7 motif, especially the central residues within the region, in hA3G dimerization, hA3G--Vif interaction, Vif-mediated hA3G degradation as well as subcellular localization of hA3G. This work highlights a multiple-task interface formed by loop7 motif, which regulates biological function of hA3G, thus providing the feasibility of the strategy of blocking Vif-mediated A3G degradation by targeting the putative site around loop7.

Recent studies find a number of promising drug targets which can be applied for increasing tumor radiosensitivity in addition to normal tissue radioprotection,including oxygen free radical scavenger,drug targets in view of DNA damage repair and cell cycle,new targets inhibiting cell death,radioprotection mediated by growth factors,regulation of the cell signaling pathways,angiotensin-converting enzyme inhibitor.radiorestorative chemicals and so on.

Artemisinin and its derivative is a kind of efficient,quick and low toxicity of anti-malarial drug.In recent years,we find that artemisinin drugs can not only against malaria,but also have pharmacological effects of immunosuppression,antiviral and anticancer.A number of studies have confirmed that artemisinin and its derivatives have the radiosensitization effects in nasopharyngeal carcinoma,cervical cancer,lung cancer and glioma,and their mechanisms are related to decreasing Weel protein expression,increasing Cyclin B1 protein expression,blocking G2-M phase and cell apoptosis.

mRNA in ARE and GW9662 group were 2.276 ±0.534 and 0.362 ±0.026,respectively.Compared with control group,PPARγmRNA level in both of ARE and GW9662 group reached statistical significance (t =4.785,P =0.001 ;t =2.395,P =0.044).PPARγprotein expression in ARE group,GW9662 +ARE group and control group were 27 688.33 ±3 593.06,21 816.00 ±1 644.07,17 716.33 ±2 273.95,respectively,which was higher in ARE group than that in control and GW+ARE group (t =5.159,P =0.001 ;t =3.038,P =0.016). NF-κB p65 mRNA expression in GW9662 +ARE group was 0.346 ±0.149,which in ARE group and GW9662 group were 0.392 ±0.1 87 and 1 .720 ±0.338,respec-tively.The differences of NF-κB p65 mRNA expression level between ARE,and control or GW9662 group were statistically significant (t =3.592,P =0.007;t =7.851 ,P =0.000).While,the differences of Caspase-3 mRNA and protein expression levels among the four groups were not statistically significant (F =1 .1 81 ,P =0.376;F =0.647,P >0.05).Conclusion ARE may restrain NF-κB through up-regulating PPARγto inhibit the proliferation and invasive potential of LLC in vitro, which suggests that PPAR-γmay be a novel therapeutic target for lung cancer.