Prevalence and time course of postoperative nausea and vomiting and severe pain in patients under general anesthesia with patient-controlled intravenous analgesia

Introduction: Postoperative nausea and vomiting (PONV) and pain are common and distressing complications in patients undergoing surgery. However, it remains uncertain whether timing of the postoperative course or the diel rhythm influences the occurrence of PONV or severe pain. Therefore, we aimed to explore the temporal distribution of PONV and severe pain. Material and methods: In this prospective observational study, we enrolled patients aged 18–65 years with American Society of Anesthesiologists classifications I–III, who were scheduled for surgery under general anesthesia. Patients were visited postoperatively at regular intervals (every 6 h over a 24-h period). Incidence of PONV was recorded and categorized based on real-time divisions: before dawn (00:00–05:59), morning (06:00–11:59), afternoon (12:00–17:59), and evening (18:00–23:59) and as sequential periods (i.e., 0–6, 6–12, 12–18, and 18–24 h). Severe pain and use of additional remedies were also recorded. Results: A total of 724 patients were included in the final analysis. Of these, 14.92 % experienced PONV within the first 6 h, and 8.29 % received antiemetic therapy. Occurrence of PONV and administration of remedies declined over the 24-h postoperative period. The lowest rate of PONV was observed during the pre-dawn hours (5.66 %). There was no statistically significant difference in the incidence of PONV 24-h postoperatively between surgeries with different end times. Patients underwent orthopedic surgeries had the highest incidence of PONV during 18:00–23:59, gynecological surgery patients had the highest incidence at 12:00–17:59, and 6:00–11:59 for other surgery patients. All patients had the lowest incidence during 0:00–5:59. During the initial 6-h postoperative period, 24.59 % of patients experienced severe pain, which declined in the remaining episodes. Patients who underwent orthopedic and gynecological surgeries exhibited similar temporal patterns and distribution characteristics of PONV and severe pain. Discussion: Both PONV and severe pain declined within the 24-h postoperative period, particularly within the first 6 h. Additionally, the onset patterns of PONV vary among patients undergoing different types of surgeries, all patients demonstrated decreased susceptibility to PONV between 00:00–05:59. Our findings enhance prevention and treatment strategies within an optimized timeframe during the postoperative course

PINK1/Parkin pathway-mediated mitophagy by AS-IV to explore the molecular mechanism of muscle cell damage

Background: Functional disorders of mitochondria are closely related to muscle diseases. Many studies have also shown that oxidative stress can stimulate the production of a large number of reactive oxygen species (ROS), which have various adverse effects on mitochondria and can damage muscle cells. Purpose: In this study, based on our previous research, we focused on the PINK1/Parkin pathway to explore the mechanism by which AS-IV alleviates muscle injury by inhibiting excessive mitophagy. Methods: L6 myoblasts were treated with AS-IV after stimulation with hydrogen peroxide (H2O2) and carbonyl cyanide m-chlorophenylhydrazone (CCCP). Then, we detected the related indices of oxidative stress and mitophagy by different methods. A PINK1 knockdown cell line was established by lentiviral infection to obtain further evidence that AS-IV reduces mitochondrial damage through PINK1/Parkin. Results: After mitochondrial damage, the expression of malondialdehyde (MDA) and intracellular ROS in L6 myoblasts significantly increased, while the expression of superoxide dismutase (SOD) and ATP decreased. The mRNA and protein expression levels of Tom20 and Tim23 were decreased, while those of VDAC1 were increased. PINK1, Parkin, and LC3 II mRNA and protein expression increased, and P62 mRNA and protein expression decreased·H2O2 combined with CCCP strongly activated the mitophagy pathway and impaired mitochondrial function. However, abnormal expression of these factors could be reversed after treatment with AS-IV, and excessive mitochondrial autophagy could also be reversed, thus restoring the regulatory function of mitochondria. However, AS-IV-adjusted function was resisted after PINK1 knockdown. Conclusion: AS-IV is a potential drug for myasthenia gravis (MG), and its treatment mechanism is related to mediating mitophagy and restoring mitochondrial function through the PINK1/Parkin pathway

TRIM21 inhibits the replication of H5N1 and H3N2 virus rather than H7N9 virus.

(A-D) TRIM21-expressing A549 cells were infected separately with H7N9, H5N1 and H3N2 at an MOI = 1.0 for 12 h, and then the levels of protein (A), mRNA (B), vRNA (C), and the TCID50 (D) were examined. Wild-type A549 cells were used as the control (*, p 0.05; **, p 0.01; ns, p > 0.05). (E-H) TRIM21-KO A549 cells were infected separately with H7N9, H5N1 and H3N2 at an MOI = 1.0 for 12 h, and then the levels of protein (E), mRNA (F), vRNA (G), and the TCID50 (H) were examined. Wild-type A549 cells were used as the control. Each experiment was independently performed with three biological repeats. All results are presented as means ± SD. *, p 0.05; **, p 0.01; ns, p > 0.05. (TIF)</p

TRIM21 directly interacts with the M1 protein of H9N2, H3N2, and H5N1 viruses.

(A) Mass spectrometry identification of TRIM21 as a potential binding partner of H9N2 M1. A549 cells infected with H9N2 virus at an MOI = 1.0 for 24 h were immunoprecipitated with anti-M1 mAb or mouse IgG, and analyzed by mass spectrometry. The TRIM21 was detected in anti-M1 IP samples. Red indicates matched B ions, blue indicates matched Y ions, green indicates unmatched ions, grey indicates precursor ions. (B) TRIM21 interacts with H9N2 M1. Myc-tagged TRIM21, Flag-GST-tagged H9N2 M1 plasmids were transfected into HEK293T cells individually. Cell lysates were subjected to coimmunoprecipitation and western blotting using the indicated antibodies. (C) Endogenous association of TRIM21 with H9N2 M1. A549 cells infected with H9N2 virus at an MOI = 1.0 for 24 h were immunoprecipitated with anti-M1 or control IgG, and analyzed by western blotting with the indicated antibodies. (D) TRIM21 interacts directly with H9N2 M1. Purified His-H9-M1 protein mixed with the GST or GST-TRIM21 proteins was pulled down with GST Sepharose, and analyzed by western blotting with the corresponding antibodies. (E) TRIM21 colocalizes with H9N2 M1. HEK293T cells were transfected with Flag-tagged vector or Flag-tagged TRIM21 plasmid for 24 h, infected with H9N2 virus at an MOI = 1.0 for 12 h, and then incubated with the anti-Flag rabbit mAb, anti-Myc mouse mAb, anti-M1 mouse mAb, FITC-Labeled goat anti-rabbit IgG, and Alexa Fluor 546-conjugated donkey anti-mouse IgG. DAPI staining revealed the nuclei. The images were obtained by confocal microscopy. Scale bar = 10 mm. (F) TRIM21 interacts with H3N2 M1 and H5N1 M1. Myc-tagged TRIM21 and Flag-GST-tagged PR8 M1 or H3N2 M1, H5N1 M1, H7N9 M1 plasmids were individually transfected into HEK293T cells. Cell lysates were subjected to coimmunoprecipitation and western blotting using the indicated antibodies. (G) Endogenous association of TRIM21 with H3N2 M1 and H5N1 M1. A549 cells infected with PR8, H3N2, H5N1 and H7N9 viruses at an MOI = 1.0 for 24 h were immunoprecipitated with anti-M1 or control IgG, and analyzed by western blotting with the indicated antibodies. Each experiment was independently performed with three biological repeats.</p

R⁹⁵ of M1 protein is critical for interacting with the PRY/SPRY domain of TRIM21.

(A) Amino acid sequence alignment of the M1 protein from various IAV, including PR8, H3N2, H5N1, H7N9, H9N2, H5Nx, and H7Nx. (B) The structural prediction of the M1 proteins from PR8, H3N2, H5N1, H7N9, and H9N2 viruses using SWISS MODEL and PyMOL software. (C-E) R95 of M1 is required for the TRIM21 interaction. HEK293T cells were separately transfected with plasmids encoding Flag-GST H3 M1, H5 M1, H9 M1 and its mutants (T37A, R95K, S242N, K242N), Myc-tagged TRIM21 plasmid for 48 h individually, and then cell lysates were incubated as indicated for a coimmunoprecipitation assay and analyzed by western blotting with the indicated antibodies. Each experiment was independently performed with three biological repeats.</p

The primers used for cloning and quantitative real-time PCR.

The primers used for cloning and quantitative real-time PCR.</p

Short-term effectiveness of single-dose intranasal spray COVID-19 vaccine against symptomatic SARS-CoV-2 Omicron infection in healthcare workers: a prospective cohort studyResearch in context

Summary: Background: The pivotal phase 3 efficacy clinical trial has demonstrated that a two-dose regimen of dNS1-RBD (Beijing Wantai Biological Pharmacy Enterprise, Beijing, China) is well-tolerated and provides wide protection against SARS-CoV-2 infection. However, the effectiveness of a single-dose regimen is still unknown. We aimed to estimate the effectiveness of one-dose of dNS1-RBD against symptomatic Omicron infections in real-world conditions. Methods: This prospective cohort study was conducted during an Omicron outbreak among healthcare workers in Xiamen, China, from December 22, 2022 to January 16, 2023. Participants chose to receive single-dose of dNS1-RBD or remain unvaccinated based on personal preference. Healthcare workers daily validated their SARS-CoV-2 infection status, using either RT-PCR or rapid antigen test. A survey questionnaire was conducted to gather information on acute symptoms from individuals infected with SARS-CoV-2. The primary outcome was the symptomatic SARS-CoV-2 infections after enrollment in the dNS1-RBD recipients or the control group among all participants and by prior COVID-19 vaccination status. Findings: On December 22, 2022, a total of 1391 eligible participants without a history of prior SARS-CoV-2 infection were enrolled. Among them, 550 received single-dose of dNS1-RBD, while 841 remained unvaccinated. In the total cohort, the range of follow-up time was 1∼26 days. During the study period, a total of 880 symptomatic SARS-CoV-2 infections were identified in the total cohort. The adjusted vaccine effectiveness against symptomatic SARS-CoV-2 infections and the infections requiring medical attention were 19.0% (95% CI: 6.7, 29.7, P = 0.004) and 59.4% (95% CI: 25.1, 78.0, P = 0.004) in the total cohort, 11.6% (95% CI: −2.4, 23.7, P = 0.100) and 55.3% (95% CI: 15.3, 76.4, P = 0.014) in the participants with inactivated COVID-19 vaccination history, as well as 87.0% (95% CI: 72.6, 93.9, P < 0.001) and 84.2% (95% CI: −41.8, 98.2, P = 0.099) in the naïve participants, respectively. Interpretation: When administered as a booster to individuals with a history of inactivated COVID-19 vaccination, a single-dose of dNS1-RBD provides protection against infections requiring medical attention at least in the short-term after vaccination. The data also showed that a single-dose of dNS1-RBD is protective against symptomatic SARS-CoV-2 infections as a primary immunization for individuals without prior exposure, but due to the limited sample size of naïve participants, further research with a larger sample size is needed to make a solid conclusion. Funding: Xiamen Science and Technology Bureau 2022 General Science and Technology Plan Project and the Bill &amp; Melinda Gates Foundation

Excel spreadsheet containing, in separate sheets, the underlying numerical data and statistical analysis for Fig panels 3A, 4B-4D, 4F-4K, 5A, 5D-5E, 5H-5J, 6A, 6D, 6F-6H, 6J, 6M, 6O-6Q, 7F, S2B-S2D, S2F-S2H, S3B-S3D, S3F-S3H, S4C-S4G, S5A-S5B, S5G-S5H, and S5J-S5L.

Excel spreadsheet containing, in separate sheets, the underlying numerical data and statistical analysis for Fig panels 3A, 4B-4D, 4F-4K, 5A, 5D-5E, 5H-5J, 6A, 6D, 6F-6H, 6J, 6M, 6O-6Q, 7F, S2B-S2D, S2F-S2H, S3B-S3D, S3F-S3H, S4C-S4G, S5A-S5B, S5G-S5H, and S5J-S5L.</p

Effect of TRIM21 knockdown and M1 mutation (R⁹⁵K and K²⁴²N) on pathogenicity in mice.

The 8-week-old C57BL/6 mice (13 per group) were intranasally infected with WT H9N2 virus, H9N2 mutant viruses R95K, K242N, or R95K/K242N at a dose of 107.0 TCID50 H9N2 virus, respectively. In another experiment, the 3-week-old C57BL/6 mice (thirteen per group) were intranasally infected with 1011.0 TCID50 of AAV6 with the TRIM21-shRNA (shTRIM21) or scrambled shRNA control (shCtrl). Four weeks after infection, the treated mice were intranasally infected with H9N2 (107.0TCID50). Three mice per group were euthanized on day 6 post-infection to check for lesions and virus replication in the lungs. The remaining mice (10 per group) were monitored until day 14. Mice with a weight loss of more than 30% of their initial body weight were euthanized and recorded as dead. (A and B) Curves of body weight (A) and survival (B) in mice (n = 10 mice) from 0 to 14 days after infection with WT H9N2 virus or H9N2 mutant viruses R95K, K242N and R95K/K242N. (C) Gross and histopathological lesions in the lung at 6 day after infection. (D) The lesion area was measured as a percentage and μm2 of the total lung area in (C). (E-H) The lung of infected mice was harvested to detect the levels of viral proteins (E), mRNA level of M gene (F), viral RNA copies (G), and the TCID50 (H). (I) Validation of shTRIM21 knockdown effect in mice. 3-week-old C57BL/6 mice were intranasally infected with shCtrl or shTRIM21 for four weeks, and TRIM21 expression in mouse lungs was detected by Western blot using the TRIM21 antibody. (J-K) Curves of body weight (J) and survival (K) in TRIM21 knocked down mice from 0 to 14 days after infection with WT H9N2 virus (n = 10 mice). (L) Gross and histopathological lesions in lungs of TRIM21 knocked down mice at 6 days after infection. (M) The lesion area was measured as a percentage and μm2 of total lung area in (L). (N-Q) The lungs of TRIM21 knocked down mice with H9N2 infection were harvested to detect the levels of viral proteins (N), mRNA level of M gene (O), viral RNA copies (P), and the TCID50 (Q). Each experiment was independently performed with three biological repeats. All results are presented as means ± SD.*, p 0.05; **, p 0.01; ns, p > 0.05. The photo by Lulu Lin.</p

Replication ability of H9N2 virus with or without the residue R⁹⁵ of M1.

(A-E) Proportions of the R95K mutation in the M1 protein of H3N2 (A), H5N1 (B), H9N2 (C), H7N9 (D), and H1N1 (E) viruses. M1 sequence data of different IAV subtypes were obtained from the NCBI GenBank Database. The image was created using the website https://app.biorender.com/. (F) A549 cells were co-infected with WT (MOI = 0.1) and R95K mutant H9N2 viruses (MOI = 0.1, 0.5. and 1.0) for 12 h, and then the vRNA was examined. Each experiment was independently performed with three biological repeats. All results are presented as means ± SD. *, p 0.05; **, p 0.01; ns, p > 0.05.</p