Inhibition of Sirt3 activates the cGAS-STING pathway to aggravate hepatocyte damage in hepatic ischemia-reperfusion injury mice.

Hepatic ischemia-reperfusion injury (IRI) typically manifests during subtotal hepatectomy and inflicts substantial damage to liver function in the perioperative period. Although the central role of cGAS-STING-mediated immune inflammation in hepatocyte damage during hepatic IRI is acknowledged, the precise regulatory mechanisms remain elusive. The current study aims to elucidate how Sirt3 inhibition activates the cGAS-STING pathway and exacerbates hepatocyte damage in hepatic IRI. We established both in vivo and in vitro models by creating hepatic IRI mice model and subjecting AML-12 hepatocyte cell lines to oxygen-glucose deprivation/reperfusion (OGD/R). Hepatic IRI compromised liver and mitochondrial function while elevating cytosolic mitochondrial DNA (mtDNA) levels in hepatocytes. Additionally, both in vivo hepatic IRI and in vitro OGD/R induced increased phosphorylation and activation of cGAS, STING, and IRF3, accompanied by heightened levels of pro-inflammatory factors, including TNF-α, IL-1ß, and type I interferon (IFN-ß). Importantly, knockdown of cGAS or STING through siRNA effectively attenuated hepatic IRI-induced inflammation and ameliorated liver function in both experimental settings, underscoring the dynamic involvement of the cGAS-STING pathway in hepatic IRI-induced inflammation. Furthermore, we observed a significant reduction in Sirt3 expression following hepatic IRI, both in vivo and in vitro. Then we generated Sirt3-deficient mice and applied Sirt3 knockdown in AML-12 hepatocytes. Notably, Sirt3 deficiency led to increased phosphorylation and activation of cGAS, STING, and IRF3, coupled with elevated TNF-α, IL-1ß, and IFN-ß levels in both in vivo and in vitro conditions. Moreover, upon silencing various downstream targets of Sirt3, such as transcription factors Sp1, Pu1, and p65, we observed that specifically knocking down p65 in AML-12 hepatocytes reduced cGAS mRNA levels. Co-immunoprecipitation assays confirmed a direct interaction between Sirt3 and p65. The absence of Sirt3 significantly increased nuclear translocation of p65 in mice, whereas Sirt3 knockdown in AML-12 hepatocytes heightened nuclear translocation of p65. ChIP-PCR assays demonstrated that Sirt3 deficiency notably enhanced the binding of p65 to two cGAS promoters, ultimately promoting cGAS transcription. Collectively, our results underscored that inhibition of Sirt3 activates the cGAS-STING pathway to aggravate hepatocyte damage by increasing cytosolic mtDNA and promoting nuclear translocation of p65 to promote cGAS transcription in hepatic IRI. These findings hold promise for potential therapeutic interventions in hepatic IRI by targeting the Sirt3-cGAS-STING axis, offering new avenues for the development of clinical strategies to mitigate liver damage during the perioperative period.

Microglial exosome miR-124-3p in hippocampus alleviates cognitive impairment induced by postoperative pain in elderly mice.

Cognitive impairment induced by postoperative pain severely deteriorates the rehabilitation outcomes in elderly patients. The present study focused on the relationship between microglial exosome miR-124-3p in hippocampus and cognitive impairment induced by postoperative pain. Cognitive impairment model induced by postoperative pain was constructed by intramedullary nail fixation after tibial fracture. Morphine intraperitoneally was carried out for postoperative analgesia. Morris water maze tests were carried out to evaluate the cognitive impairment, while mRNA levels of neurotrophic factors (BDNF, NG) and neurodegenerative biomarker (VILIP-1) in hippocampus were tested by q-PCR. Transmission electron microscope was used to observe the axon degeneration in hippocampus. The levels of pro-inflammatory factors (TNF-α, IL-1ß, IL-6), the levels of anti-inflammatory factors (Ym, Arg-1, IL-10) and microglia proliferation marker cyclin D1 in hippocampus were measured to evaluate microglia polarization. Bioinformatics analysis was conducted to identify key exosomes while BV-2 microglia overexpressing exosome miR-124-3p was constructed to observe microglia polarization in vitro experiments. Exogenous miR-124-3p-loaded exosomes were injected into hippocampus in vivo. Postoperative pain induced by intramedullary fixation after tibial fracture was confirmed by decreased mechanical and thermal pain thresholds. Postoperative pain induced cognitive impairment, promoted axon demyelination, decreased BDNF, NG and increased VILIP-1 expressions in hippocampus. Postoperative pain also increased pro-inflammatory factors, cyclin D1 and decreased anti-inflammatory factors in hippocampus. However, these changes were all reversed by morphine analgesia. Bioinformatics analysis identified the critical role of exosome miR-124-3p in cognitive impairment, which was confirmed to be down-regulated in hippocampus of postoperative pain mice. BV-2 microglia overexpressing exosome miR-124-3p showed decreased pro-inflammatory factors, cyclin D1 and increased anti-inflammatory factors. In vivo, stereotactic injection of exogenous miR-124-3p into hippocampus decreased pro-inflammatory factors, cyclin D1 and increased anti-inflammatory factors. The cognitive impairment, axon demyelination, decreased BDNF, NG and increased VILIP-1 expressions in hippocampus were all alleviated by exogenous exosome miR-124-3p. Microglial exosome miR-124-3p in hippocampus alleviates cognitive impairment induced by postoperative pain through microglia polarization in elderly mice.

Ischemic preconditioning attenuates endoplasmic reticulum stress-dependent apoptosis of hepatocytes by regulating autophagy in hepatic ischemia-reperfusion injury.

Hepatic ischemia-reperfusion injury (HIRI) usually occurs during subtotal hepatectomy and severely damages liver function during the perioperative period. Endoplasmic reticulum stress (ERS) dependent apoptosis has been suggested to play a crucial role in HIRI progression. The present study focused on the regulatory effect of autophagy activation induced by ischemic preconditioning (IPC) on ERS-dependent apoptosis of hepatocytes in HIRI. A HIRI mouse model and oxygen-glucose deprivation/reperfusion (OGD/R) AML-12 hepatocyte cell lines were constructed to evaluate the protective effect of IPC in vivo and in vitro. The protein levels of p-eIF2α, CHOP, and cleaved caspase-12 were used to evaluate the ERS-dependent apoptosis, whereas LC3-II and p62 were considered as the autophagy activation markers. The beneficial molecular chaperones GRP78, HSP60, and HSP70 were also tested to evaluate autophagy. HIRI significantly increased ERS-dependent apoptosis markers and the number of apoptotic cells and damaged liver function. The ERS inhibitor salubrinal significantly alleviated liver injury in HIRI and OGD/R hepatocytes. Furthermore, both remote IPC and direct IPC significantly alleviated liver injury and inflammatory cell infiltration. IPC also upregulated LC3-II, downregulated p62 expression, and increased the mRNA levels of GRP78, HSP60, and HSP70 in HIRI mice and OGD/R hepatocytes, indicating the activation of autophagy by IPC. The autophagy inhibitor 3-methyladenine significantly attenuated the protective effects of IPC on ERS-dependent apoptosis and liver function, whereas the autophagy activator rapamycin mimicked the protective effects of IPC on ERS-dependent apoptosis in vivo and in vitro, suggesting a regulatory role of autophagy in ERS-dependent apoptosis. These results demonstrated that IPC could induce moderate autophagy and upregulate a few molecular chaperones to strengthen endogenous defense mechanisms, which is beneficial for alleviating ERS-dependent apoptosis and protecting hepatocytes from HIRI.

Novel approach by natural language processing for COVID-19 knowledge discovery.

BACKGROUND: The impact of COVID-19 on public health has mandated an 'all hands on deck' scientific response. The current clinical study and basic research on COVID-19 are mainly based on existing publications or our knowledge of coronavirus. However, efficiently retrieval of accurate, relevant knowledge on COVID-19 can pose significant challenges for researchers. METHODS: To improve quality in accessing important literature findings, we developed a novel natural language processing (NLP) method to automatically recognize the associations among potential targeted host organ systems, associated clinical manifestations, and pathways. We further validated these associations through clinician experts' evaluations and prioritize candidate drug targets through bioinformatics network analysis. RESULTS: We found that the angiotensin-converting enzyme 2 (ACE2), a receptor that SARS-CoV-2 required for cell entry, is associated with cardiovascular and endocrine organ system and diseases. Furthermore, we found SARS-CoV-2 is associated with some important pathways such as IL-6, TNF-alpha, and IL-1 beta-induced dyslipidemia, which are related to inflammation, lipogenesis, and oxidative stress mechanisms, suggesting potential drug candidates. CONCLUSION: We prioritized the list of therapeutic targets involved in antiviral and immune modulating drugs for experimental validation, rendering it valuable during public health crises marked by stresses on clinical and research capacity. Our automatic intelligence pipeline also contributes to other novel and emerging disease management and treatments in the future.

Post-traumatic stress disorder in patients with rheumatic disease during the COVID-19 outbreak: a cross-sectional case-control study in China.

OBJECTIVE: The COVID-19 pandemic is not only a traumatic event, but a collective stressor unfolding over time, causing devastating implications for the mental health. This study aimed to shed light on the mental health status of patients with rheumatic disease (RD) during the massive outbreak of COVID-19 in China, especially the prevalence and severity of post-traumatic stress disorder (PTSD) compared with healthy individuals. METHODS: A total of 486 patients with RD and 486 age-matched and sex-matched healthy individuals were recruited into the study. For each participant, we collected demographic and clinical characteristics data. The PTSD Checklist for DSM-5 (PCL-5) and four items from the Pittsburgh Sleep Quality Index (PSQI) were used to investigate the prevalence and severity of PTSD and sleep quality, respectively. RESULTS: Compared with healthy control subjects (n=486), patients with RD (n=486) had a higher prevalence of PTSD (12.1% vs 4.1%; p<0.001). Higher total scores on the PCL-5 and on all four items from the PSQI (p≤0.001) were also observed. Female, old age, poor sleep quality, long duration of RD, poor subjective evaluation of the disease and pessimistic subjective perception of the epidemic were identified as risk factors of PTSD in patients with RD during the COVID-19 epidemic. CONCLUSION: During the COVID-19 outbreak, patients with RD presented a higher prevalence and severity of PTSD and showed more sleep disturbances. Our findings confirm the importance of psychological assessment and mental healthcare out of regular clinical care for patients with RD during the pandemic.

Antiviral agents, glucocorticoids, antibiotics, and intravenous immunoglobulin in 1142 patients with coronavirus disease 2019: a systematic review and meta-analysis.

INTRODUCTION: The treatment effects of antiviral agents, glucocorticoids, antibiotics, and intravenous immunoglobulin are controversial in patients with coronavirus disease 2019 (COVID­19). OBJECTIVES: This study aimed to evaluate the impact of drug therapy on the risk of death in patients with COVID­19. PATIENTS AND METHODS: The PubMed, Embase, Web of Science, Cochrane Library, and major preprint platforms were searched to retrieve articles published until April 7, 2020. Subsequently, the effects of specific drug interventions on mortality of patients with COVID­19 were assessed. Odds ratios (ORs) and relative risks (RRs) with corresponding 95% CIs were pooled using random effects models. RESULTS: Of 3421 references, 6 studies were included. Pooled results from retrospective studies revealed that antiviral agents may contribute to survival benefit (OR, 0.42; 95% CI, 0.17-0.99; P = 0.048; I2 = 82.8%), whereas a single randomized controlled trial found no effects of an antiviral agent on mortality (RR, 0.77; 95% CI, 0.45-1.3; P = 0.33). Glucocorticoid use led to an increased risk of death (OR, 2.43; 95% CI, 1.44-4.1; P = 0.001; I2 = 61.9%). Antibiotics did not significantly affect mortality (OR, 1.13; 95% CI, 0.67-1.89; P = 0.64; I2 = 0%). Similarly, intravenous immunoglobulin had a nonsignificant effect on mortality (OR, 2.66; 95% CI, 0.72-9.89; P = 0.14; I2 = 93.1%). CONCLUSIONS: With the varied heterogeneities across interventions, the current evidence indicated a probable survival benefit from antiviral agent use and a harmful effect of glucocorticoids in patients with COVID­19. Neither any of antibiotics nor intravenous immunoglobulin were associated with survival benefit in this population.