DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition.

INTRODUCTION: Intrauterine adhesions (IUA) manifest as endometrial fibrosis, often causing infertility or recurrent miscarriage; however, their pathogenesis remains unclear. OBJECTIVES: This study assessed the role of Dickkopf WNT signaling pathway inhibitor 1 (DKK1) and autophagy in endometrial fibrosis, using clinical samples as well as in vitro and in vivo experiments. METHODS: Immunohistochemistry, immunofluorescence and western blot were used to determine the localization and expression of DKK1 in endometrium; DKK1 silencing and DKK1 overexpression were used to detect the biological effects of DKK1 silencing or expression in endometrial cells; DKK1 gene knockout mice were used to observe the phenotypes caused by DKK1 gene knockout. RESULTS: In patients with IUA, DKK1 and autophagy markers were down-regulated; also, α-SMA and macrophage localization were increased in the endometrium. DKK1 conditional knockout (CKO) mice showed a fibrotic phenotype with decreased autophagy and increased localization of α-SMA and macrophages in the endometrium. In vitro studies showed that DKK1 knockout (KO) suppressed the autophagic flux of endometrial stromal cells. In contrast, ectopic expression of DKK1 showed the opposite phenotype. Mechanistically, we discovered that DKK1 regulates autophagic flux through Wnt/ß-catenin and PI3K/AKT/mTOR pathways. Further studies showed that DKK1 KO promoted the secretion of interleukin (IL)-8 in exosomes, thereby promoting macrophage proliferation and metastasis. Also, in DKK1 CKO mice, treatment with autophagy activator rapamycin partially restored the endometrial fibrosis phenotype. CONCLUSION: Our findings indicated that DKK1 was a potential diagnostic marker or therapeutic target for IUA.

Olanzapine for the prevention of postoperative nausea and vomiting after gynecologic laparoscopic surgery: a randomized controlled trial.

Purpose: This study was designed to investigate the prophylactic effect of oral olanzapine in postoperative nausea and vomiting after gynecologic laparoscopic surgery. Methods: ASA I-II, aged 18-75 years, planned to undergo gynecologic laparoscopic surgery with general anesthesia in adult female patients. Using the randomized numbers table, the patients were placed in two groups. Oral olanzapine 5 mg or placebo was given 1 h before anesthesia. All patients received standard antiemetic prophylaxis with dexamethasone and granisetron. The primary outcome was nausea and/or vomiting in the 24 h after the postoperative. Results: A total of 250 patients were randomized, and 241 were analyzed. The primary outcome occurred in 10 of 120 patients (8.3%) in the olanzapine group and 23 of 121 patients (19.2%) in the placebo group (p = 0.014). According to Kaplan-Meier analysis, the probabilities of nausea and/or vomiting in the 24 h after the postoperative in the olanzapine group were lower than in the placebo group (log-rank p = 0.014). In a multivariate Cox analysis, the variables of use of olanzapine [hazard ratio (HR): 0.35, 95% confidence interval (CI): 0.16-0.79; p = 0.012] and use of vasoactive drugs (HR: 2.48, 95% CI: 1.07-5.75; p = 0.034) were independently associated with nausea and/or vomiting in the 24 h after the postoperative. Conclusion: Our data suggest that olanzapine relative to placebo decreased the risk of nausea and/or vomiting in the 24 h after gynecologic laparoscopic surgery. Trial registration: The trial was registered prior to patient enrollment at The Chinese Clinical Trial Registry (https://www.chictr.org.cn/showproj.html?proj=166900, link to registry page, Principal investigator: Nanjin Chen, Date of registration: 25 April 2022).

Preventing nausea and vomiting after laparoscopic gynecological surgery: the benefits of using olanzapine Why was this study done? Despite the use of antiemetics, postoperative nausea and vomiting remain prevalent. Furthermore, patients who undergo gynecological laparoscopic surgery are at an increased risk. Therefore, this study investigated whether oral Olanzapine could reduce the incidence of nausea and vomiting after gynaecological Laparoscopy? What did the researchers do? The research team examined patients who underwent gynecological laparoscopic surgery under general anesthesia. They observed the occurrence of nausea and vomiting within 24 hours after surgery in patients who either received or did not receive Olanzapine treatment. The goal was to assess the effectiveness of Olanzapine in reducing postoperative nausea and vomiting. What did the researchers find? The addition of Olanzapine, when combined with granisetron and dexamethasone, resulted in a decreased risk of nausea and/or vomiting within the 24 hours following gynecologic laparoscopic surgery, as compared to the placebo. Administering oral Olanzapine at a dosage of 5 mg reduced the incidence of nausea and vomiting after gynecological laparoscopy from 19.2% to 8.3%. What do the findings mean? This study has identified a safe and effective medication for preventing postoperative nausea and vomiting. Implementing Olanzapine as a preventive measure can significantly reduce the incidence of nausea and vomiting following surgery, thereby enhancing the overall medical experience for patients.

Nocturnal oxygen saturation is associated with all-cause mortality: a community-based study.

STUDY OBJECTIVES: Observational studies have demonstrated the association between the single-point measurement of oxygen saturation (SpO2) level and mortality in the general population. This study aimed to evaluate whether nocturnal SpO2 level could predict all-cause mortality in a community-based population. METHODS: The study samples were obtained from the Sleep Heart Health Study, which included 2,280 men and 2,606 women (mean age, 63.8 ± 11.1 years). A pulse oximeter based on overnight in-home polysomnography was used to monitor SpO2 levels during total sleep time (SpO2-TOTAL). Multivariable Cox proportional hazards analysis was performed to examine the association between nocturnal SpO2 and all-cause mortality. RESULTS: During the follow-up period of 10.7 ± 3.0 years, 1,110 (22.7%) people died. After adjusting for confounding factors, multivariable Cox regression analysis showed that the average SpO2-TOTAL (hazard ratio [HR] 0.93; 95% confidence interval [CI] 0.90-0.96, P < .001) was associated with all-cause mortality. These findings remained stable in individuals with low and high apnea-hypopnea index levels. Additionally, maximum SpO2-TOTAL (HR, 0.91; 95% CI, 0.87-0.96; P = .001) and minimum SpO2-TOTAL (HR, 0.98; 95% CI, 0.97-0.99; P = .001) could predict all-cause mortality. A significant association between nocturnal hypoxemia and all-cause mortality was also observed. CONCLUSIONS: Our findings highlight the importance of monitoring nocturnal SpO2 level and improving hypoxemia in the general populations. CITATION: Yan B, Gao Y, Zhang Z, Shi T, Chen Q. Nocturnal oxygen saturation is associated with all-cause mortality: a community-based study. J Clin Sleep Med. 2024;20(2):229-235.

GWAS and Meta-QTL Analysis of Yield-Related Ear Traits in Maize.

Maize ear traits are an important component of yield, and the genetic basis of ear traits facilitates further yield improvement. In this study, a panel of 580 maize inbred lines were used as the study material, eight ear-related traits were measured through three years of planting, and whole genome sequencing was performed using the maize 40 K breeding chip based on genotyping by targeted sequencing (GBTS) technology. Five models were used to conduct a genome-wide association study (GWAS) on best linear unbiased estimate (BLUE) of ear traits to find the best model. The FarmCPU (Fixed and random model Circulating Probability Unification) model was the best model for this study; a total of 104 significant single nucleotide polymorphisms (SNPs) were detected, and 10 co-location SNPs were detected simultaneously in more than two environments. Through gene function annotation and prediction, a total of nine genes were identified as potentially associated with ear traits. Moreover, a total of 760 quantitative trait loci (QTL) associated with yield-related traits reported in 37 different articles were collected. Using the collected 760 QTL for meta-QTL analysis, a total of 41 MQTL (meta-QTL) associated with yield-related traits were identified, and 19 MQTL detected yield-related ear trait functional genes and candidate genes that have been reported in maize. Five significant SNPs detected by GWAS were located within these MQTL intervals, and another three significant SNPs were close to MQTL (less than 1 Mb). The results provide a theoretical reference for the analysis of the genetic basis of ear-related traits and the improvement of maize yield.

A rapid multispectral endoscopic imaging system for in vivo assessment of the morphological and physiological characteristics of mouse intestines.

Accurate assessment of blood content in biological tissues is critical for the diagnosis and monitoring of various diseases, including cardiovascular disease, tumors, trauma, and the success rate of organ transplants. In this study, a multispectral endoscopic imaging system was built for capturing tissue reflection optical images in 18 bands across the wavelength range from 400 nm to 760 nm, non-invasively. The system was characterized by six tri-channel narrowband filters installed in front of the light source to achieve spectral separation and was equipped with a specially designed color CCD for achieving a speed of 24 multispectral imaging cubes per second. A method based on linear matrix inversion was proposed to calibrate the CCD spectral response overlaps, while a spectral analysis algorithm was developed for evaluating blood content and detecting tissue composition. The developed system was implemented in an in vivo mouse model for illustrating the blood volume, blood oxygen saturation index, and scattering particle size of the intestinal wall mucosa. The observations not only helped us to understand the blood supply situation in the intestinal mucosa, but also further testified the feasibility of our presented system. Meanwhile, the developed system could provide critical non-invasive optical information for intracavitary cancer diagnosis, surgery guidance, and treatment assessment.

Non-destructive diagnostic testing of cardiac myxoma by serum confocal Raman microspectroscopy combined with multivariate analysis.

The symptoms of cardiac myxoma (CM) mainly occur when the tumor is growing, and the diagnosis is determined by clinical presentation. Unfortunately, there is no evidence that specific blood tests are useful in CM diagnosis. Raman spectroscopy (RS) has emerged as a promising auxiliary diagnostic tool because of its ability to simultaneously detect multiple molecular features without labelling. The objective of this study was to identify spectral markers for CM, one of the most common benign cardiac tumors with insidious onset and rapid progression. In this study, a preliminary analysis was conducted based on serum Raman spectra to obtain the spectral differences between CM patients (CM group) and healthy control subjects (normal group). Principal component analysis-linear discriminant analysis (PCA-LDA) was constructed to highlight the differences in the distribution of biochemical components among the groups according to the obtained spectral information. Principal component analysis was combined with a support vector machine model (PCA-SVM) based on three different kernel functions (linear, polynomial, and Gaussian radial basis function (RBF)) to resolve spectral variations between all study groups. The results showed that CM patients had lower serum levels of phenylalanine and carotenoid than those in the normal group, and increased levels of fatty acids. The resulting Raman data was used in a multivariate analysis to determine the Raman range that could be used for CM diagnosis. Also, the chemical interpretation of the spectral results obtained is further presented in the discussion section based on the multivariate curve resolution-alternating least squares (MCR-ALS) method. These results suggest that RS can be used as an adjunct and promising tool for CM diagnosis, and that vibrations in the fingerprint region can be used as spectral markers for the disease under study.

Raman microspectroscopy based TNM staging and grading of breast cancer.

The tumor-node-metastasis (TNM) system is the most common way that doctors determine the anatomical extent of cancer on the basis of clinical and pathological criteria. In this study, a spectral histopathological study has been carried out to bridge Raman micro spectroscopy with the breast cancer TNM system. A total of seventy breast tissue samples, including healthy tissue, early, middle, and advanced cancer, were investigated to provide detailed insights into compositional and structural variations that accompany breast malignant evolution. After evaluating the main spectral variations in all tissue types, the generalized discriminant analysis (GDA) pathological diagnostic model was established to discriminate the TNM staging and grading information. Moreover, micro-Raman images were reconstructed by K-means clustering analysis (KCA) for visualizing the lobular acinar in healthy tissue and ductal structures in all early, middle and advanced breast cancer tissue groups. While, univariate imaging techniques were adapted to describe the distribution differences of biochemical components such as tryptophan, ß-carotene, proteins, and lipids in the scanned regions. The achieved spectral histopathological results not only established a spectra-structure correlations via tissue biochemical profiles but also provided important data and discriminative model references for in vivo Raman-based breast cancer diagnosis.

Triptolide Alleviates Neuropathic Pain by Inhibiting the Activation of Microglial Toll-Like Receptor 3.

BACKGROUND: Current data indicates the incidence of neuropathic pain after surgical nerve injury is as high as 50%, thus representing a major problem for patients and for the medical system. Triptolide, a traditional Chinese herb, has anti-inflammatory effects on various neurodegenerative and neuroinflammatory diseases. This agent also reduces peripheral nerve injury-induced neuropathic pain, although the mechanism underlying this effect is still unknown. MATERIALS AND METHODS: The effects of triptolide on spinal nerve ligation (SNL) injury-induced neuropathic pain was studied in an animal model using behavioral, morphological and molecular biological methods. RESULTS: Repeated administration of intrathecal triptolide was found to alleviate SNL- or Poly(I:C) (toll-like receptor 3 agonist) injection-induced mechanical allodynia without any motor impairment. The mechanism by which triptolide reduces SNL- and Poly(I:C) injection-induced microglial activation appears to be via the inhibition of OX42 expression, which is a microglial-specific marker. Intrathecal triptolide also suppressed SNL- and Poly(I:C) injection-induced expression of spinal TRIF. TRIF transmits signals from activated TLR3 and is the downstream adaptor of TLR3 in microglia. In addition, intrathecal triptolide inhibited the expression of spinal pro-inflammatory IL-1 ß following SNL or Poly(I:C) injection. CONCLUSIONS: Intrathecal triptolide can suppress the TLR3/TRIF/IL-1 ß pathway in spinal microglia following SNL. This could be the underlying mechanism by which triptolide alleviate neuropathic pain induced by peripheral nerve injury.

Effect of visceral adipose tissue mass on coronary artery disease and heart failure: A Mendelian randomization study.

OBJECTIVES: Obesity is associated with coronary artery disease (CAD) and its risk factors in observational studies. This two-sample Mendelian randomization (MR) study investigated the effect of visceral adipose tissue (VAT) mass on coronary artery disease (CAD), myocardial infarction (MI) and heart failure (HF). METHODS: Genetic variants (220 SNPs, P < 5 × 10-8) associated with VAT mass were obtained from a genome-wide association study (GWAS) in the UK Biobank. Genetic associations with CAD outcomes including CAD and myocardial infarction (MI) were obtained from the CARDIoGRAMplusC4D 1000 Genomes-based GWAS (including up to 60,801 cases and 123,504 controls) and data on patients with HF were obtained from the HERMES Consortium (47,309 cases and 930,014 controls). The effects of VAT mass on CAD outcomes and HF were estimated using the inverse variance weighted (IVW) method. Sensitivity analysis and multi-variable MR were used to examine the stability of the IVW results. RESULTS: Our results showed that genetically predicted higher VAT mass was associated with increased risk of CAD (odds ratio [OR] 1.57, 95% confidence interval [CI], 1.44-1.71; P = 7.62 × 10-24), MI (OR 1.63, 95% CI: 1.48-1.79; P = 3.76 × 10-24) and HF (OR 1.71, 95% CI: 1.60-1.83; P = 1.72 × 10-53). These findings remained significant in the sensitivity analysis and multi-variable MR. CONCLUSION: This MR study suggests that genetic determinants of VAT mass are causally associated with CAD, MI and HF.

Epistasis Activation Contributes Substantially to Heterosis in Temperate by Tropical Maize Hybrids.

Epistasis strongly affects the performance of superior maize hybrids. In this study, a multiple-hybrid population, consisting of three hybrid maize sets with varied interparental divergence, was generated by crossing 28 temperate and 23 tropical inbred lines with diverse genetic backgrounds. We obtained 1,154 tested hybrids. Among these tested hybrids, heterosis increased steadily as the heterotic genetic distance increased. Mid-parent heterosis was significantly higher in the temperate by tropical hybrids than in the temperate by temperate hybrids. Genome-wide prediction and association mapping was performed for grain weight per plant (GWPP) and days to silking (DTS) using 20K high-quality SNPs, showing that epistatic effects played a more prominent role than dominance effects in temperate by tropical maize hybrids. A total of 33 and 420 epistatic QTL were identified for GWPP and DTS, respectively, in the temperate by tropical hybrids. Protein-protein interaction network and gene-set enrichment analyses showed that epistatic genes were involved in protein interactions, which play an important role in photosynthesis, biological transcription pathways, and protein synthesis. We showed that the interaction of many minor-effect genes in the hybrids could activate the transcription activators of epistatic genes, resulting in a cascade of amplified yield heterosis. The multiple-hybrid population design enhanced our understanding of heterosis in maize, providing an insight into the acceleration of hybrid maize breeding by activating epistatic effects.

CircRAB11FIP1 promoted autophagy flux of ovarian cancer through DSC1 and miR-129.

At present, no systematic and in-depth study is available on the function and potential mechanisms of circular RNA in autophagy. This study aimed to screen the expression profiles of circRNA, miRNA, and mRNA of ovarian cancer cells induced by Torin 1 (10 µM). The expression profiles of circRNA, miRNA, and mRNA were analyzed with next-generation sequencing technology. CircRAB11FIP1 expression was elevated in epithelial ovarian cancer (EOC) tissues than in normal ovarian tissues. Silencing circRAB11FIP1 inhibited the autophagic flux of ovarian cancer SKOV3 cells. However, circRAB11FIP1 overexpression activated the autophagic flux of ovarian cancer A2780 cells. CircRAB11FIP1-induced autophagy accelerated EOC proliferation and invasion. Also, circRAB11FIP1 directly bound to miR-129 and regulated its targets ATG7 and ATG14. CircRAB11FIP1 bound to desmocollin 1to facilitate its interaction with ATG101. Also, circRAB11FIP1 directly bound to the mRNA of fat mass and obesity-associated protein and promoted its expression. Then, circRAB11FIP1 mediated mRNA expression levels of ATG5 and ATG7 depending on m6A. In general, this study demonstrated that circRAB11FIP1 regulated ATG7 and ATG14 by sponging miR-129. The data suggested that circRAB11FIP1 might serve as a candidate biomarker for EOC diagnosis and treatment.

Early label-free analysis of mitochondrial redox states by Raman spectroscopy predicts septic outcomes.

BACKGROUND: Sepsis remains an unacceptably high mortality due to the lack of biomarkers for predicting septic outcomes in the early period. Mitochondrial redox states play a pivotal role in this condition and are disturbed early in the development of sepsis. Here, we hypothesized that visualizing mitochondrial redox states via resonance Raman spectroscopy (RRS) could identify septic outcomes at an early time point. Sepsis was induced by cecal ligation and puncture (CLP). We applied RRS analysis at baseline and 30 min, 1 h, 2 h, 4 h, and 6 h after CLP, and the mitochondrial redox states were identified. The levels of blood lactate as a predictor in sepsis were assessed. Our study is the first to reveal the possibility of in vivo detection of the mitochondrial redox state by using RRS in septic mice. The peak area for the Raman reduced mitochondrial fraction, the indicator of mitochondrial redox states, fluctuated significantly at 2 h after CLP. This fluctuation occurred earlier than the change in lactate level. Moreover, this fluctuation had higher prognostic accuracy for mortality than the lactate level during sepsis and could be a novel diagnostic marker for predicting septic outcomes according to the cutoff value of 1.059, which had a sensitivity of 80% and a specificity of 90%. OBJECTIVES: To explore an effective indicator concerning mitochondrial redox states in the early stage of sepsis and to predict septic outcomes accurately in vivo using non-invasive and label-free Resonance Raman spectroscopy (RRS) analysis. METHODS: Mitochondria, primary skeletal muscle cells andex-vivo muscles harvested from gastrocnemius were detected mitochondrial redox states respectively by using RRS. Sepsis was induced by cecal ligation and puncture (CLP). We applied RRS analysis at baseline and 30 min, 1 h, 2 h, 4 h, and 6 h after CLP, and the mitochondrial redox states were identified. The levels of blood lactate as a predictor in sepsis were assessed. The predictive correlation of mitochondrial redox states on mortality, inflammation and organ dysfunction was further assessed. RESULTS: Mitochondrial redox states were clearly recognized in ex-vivo gastrocnemius muscles as well as purified mitochondria and primary skeletal muscle cells by using RRS. The peak area for the Raman reduced mitochondrial fraction, the indicator of mitochondrial redox states, fluctuated significantly at 2 h after CLP. This fluctuation occurred earlier than the change in lactate level. Moreover, this fluctuation had higher prognostic accuracy for mortality than the lactate level during sepsis and could be a novel diagnostic marker for predicting septic outcomes according to the cutoff value of 1.059, which had a sensitivity of 80% and a specificity of 90%. CONCLUSIONS: This study demonstrated that monitoring mitochondrial redox states using RRS as early as 2 h could indicate outcomes in septic mice. These data may contribute to developing a non-invasive clinical device concerning mitochondrial redox states by using bedside-RRS.

Low-dose alcohol ameliorated high fat diet-induced anxiety-related behavior via enhancing adiponectin expression and activating the Nrf2 pathway.

Long-term high-fat-diet (HFD)-induced obesity is associated with many comorbidities, such as cognitive impairment and anxiety, which are increasing public health burdens that have gained prevalence in adolescents. Although low-dose alcohol could attenuate the risk of cardiovascular disease, its mechanism on HFD-induced anxiety-related behavior remains not clear. The mice were divided into 4 groups, Control (Con), Alcohol (Alc), HFD and HFD + Alc groups. To verify the effects of low-dose alcohol on HFD-induced anxiety-related behavior, the mice were fed with HFD for 16 weeks. At the beginning of week 13, the HFD-fed mice were administered intragastrically with low-dose alcohol (0.8 g kg-1) for 4 weeks. After 4 weeks of oral administration, low-dose alcohol decreased body weight and Lee's index in HFD-induced obese mice. Moreover, low-dose alcohol alleviated the anxiety-related behaviors of obese mice in the open field test and the elevated plus maze test. The HFD-induced damage to the hippocampus was improved in hematoxylin-eosin staining assay in mice. In addition, low-dose alcohol also suppressed HFD-induced oxidative stress and increased HFD-suppressed adiponectin (APN) expression and nuclear factor erythroid 2-related factor 2 (Nrf2) activation in the hippocampus. Taken together, low-dose alcohol significantly ameliorates HFD-induced obesity, oxidative stress and anxiety-related behavior in mice, which might be related to APN upregulation, Nrf2 activation and related antioxidase expression including SOD1, HO-1, and catalase.

Effect of the COVID-19 pandemic on complications and mortality of patients with cardiac surgery.

OBJECTIVES: The purpose of this study was to assess changes in cardiovascular disease severity, types, postoperative complications and prognosis during the COVID-19 pandemic and to explore possible influencing factors. METHODS: A total of 422 patients were enrolled in this study, and hospitalization and short-term follow-up data were retained. The patient population included 273 men and 149 women. Patients had a median (IQR) age of 54 (45-62) years and were divided into an observation group (130) and a control group (292), primarily according to severity of disease, disease types, baseline indexes, biochemical indexes, cardiac function indexes, complications and prognosis. RESULTS: During the COVID-19 pandemic, compared with the same period last year, there was a significant increase in patients with aortic dissection (27.69% vs 5.82%), a significant decrease in patients with valvular heart disease (43.08% vs 66.78%), and significantly increased emergency admission (50.00% vs 21.23%) and severity (54.62% vs 27.40%). Family company (76.37% vs 64.62%) was decreased, EuroSCORE [6.5 (2-9) vs 2 (0-5)] score, Pro-BNP [857.50 (241.00-2222.50) vs 542.40 (113.45-1776.75)] ng/L, six months mortality rate (18.46% vs 8.90%), and postoperative complications, including infected patients, atelectasis, pulmonary edema, and so on were increased, with longer length of stay in the ICU and hospital in COVID-19 pandemic. Survival analysis curve further demonstrated that it had an impact on the deaths of patients during the COVID-19 pandemic period. Through ROC analysis of the death factors of patients, it was concluded that Family company affected the death of patients, and the area under the curve was 0.654 (P < 0.05). CONCLUSIONS: In this study, we found that the admission rate of critically ill patients with cardiovascular disease, complications of cardiac surgery, and short-term mortality of patients all exhibited a short-term increase, family company may be a risk factors for short-term mortality, that may be related to public pressure caused by the COVID-19 pandemic.

Muscle-Derived Mitochondrial Transplantation Reduces Inflammation, Enhances Bacterial Clearance, and Improves Survival in Sepsis.

BACKGROUND: Mitochondrial transplantation is a promising strategy for the treatment of several diseases. However, the effects of mitochondrial transplantation on the outcome of polymicrobial sepsis remain unclear. METHODS: The distribution of transplanted mitochondria in cecal ligation and puncture (CLP)-operated mice was detected at 2 and 12 h after intravenous injection in the tail (n = 3). Then, the effects of mitochondrial transplantation on bacterial clearance (n = 7), systemic inflammation (n = 10), organ injury (n = 8), and mortality (n = 19) during CLP-induced sepsis were explored. Microarray analysis (n = 3) was used to testify the molecular changes associated with decreased systemic inflammation and multiorgan dysfunction in sepsis. RESULTS: The extraneous mitochondria were distributed in the lung, liver, kidney, and brain of CLP-operated mice at 2 and 12 h after intravenous injection in the tail. Mitochondrial transplantation increased the survival rate of septic mice, which was associated with decreased bacterial burden, systemic inflammation, and organ injury. Spleen samples were utilized for microarray analysis. Pathway analysis revealed that in polymicrobial sepsis, gene expression was significantly changed in processes related to inflammatory response, complement and coagulation cascades, and rejection reaction. CONCLUSIONS: These data displayed that mitochondrial replenishment reduces systemic inflammation and organ injury, enhances bacterial clearance, and improves the survival rate in sepsis. Thus, extraneous mitochondrial replenishment may be an effective adjunctive treatment to reduce sepsis-related mortality.

Mitochondrial Transplantation Attenuates Brain Dysfunction in Sepsis by Driving Microglial M2 Polarization.

Activation of microglia and mitochondrial dysfunction are two major contributors to the pathogenesis of sepsis-associated brain dysfunction. Mitochondrial dysfunction can alter the immunological profile of microglia favoring to a pro-inflammatory phenotype. Mitochondrial transplantation, as an emerging mitochondria-targeted therapy, possesses considerable therapeutic potential in various central nervous system injuries or diseases. However, the effects of mitochondrial transplantation on microglial polarization and neuroprotection after sepsis remain unclear. In this study, lipopolysaccharide (LPS)/interferon-γ (IFN-γ) and interleukin-4 (IL-4)/interleukin-13 (IL-13) were used to induce different phenotypes of BV2 microglial cells. We observed that mitochondrial content and function were enhanced in IL-4-/IL-13-stimulated microglia. In vitro, mitochondria treatment conferred neuroprotection by enhancing microglial polarization from the M1 phenotype to the M2 phenotype and suppressing microglial-derived inflammatory cytokine release. Furthermore, microglial phenotypes and behavior tests were assessed after mice were subjected to sepsis by cecal ligation and puncture (CLP) followed by intracerebroventricular injection of exogenous functional mitochondria. We found that mitochondrial transplantation induced microglial M2 rather than M1 response 24 h after sepsis. Mitochondrial transplantation improved behavioral deficits by increasing the latency time in inhibitory avoidance test and decreasing the number of crossing and rearing in the test session of open field test 10 days after CLP onset. These findings indicate that mitochondrial transplantation promotes the phenotypic conversion of microglia and improves cognitive impairment in sepsis survivors, supporting the potential use of exogenous mitochondrial transplantation therapy that may be a potential therapeutic opportunity for sepsis-associated brain dysfunction.

Electroacupuncture attenuated cerebral ischemic injury and neuroinflammation through α7nAChR-mediated inhibition of NLRP3 inflammasome in stroke rats.

BACKGROUND: Our previous research confirmed that electroacupuncture (EA) stimulus elicits neuroprotective effects against cerebral ischemic injury through α7 nicotinic acetylcholine receptor (α7nAChR)-mediated inhibition of high-mobility group box 1 release mechanism. This study investigated whether the signal transducer of α7nAChR and inhibition of NLRP3 inflammasome are involved in the neuroprotective effects of EA stimulus. METHODS: In adult male Sprague-Dawley rats, the focal cerebral ischemic injury was induced by middle cerebral artery occlusion (MCAO) models for 1.5 h. The expression of NLRP3 inflammasome in the penumbral tissue following reperfusion was assessed by western blotting and immunoflourescent staining. The infarct size, neurological deficit score, TUNEL staining and the expression of proinflammatory factors or anti-inflammatory cytokines were evaluated at 72 h after reperfusion in the presence or absence of either α7nAChR antagonist (α-BGT) or agonist (PHA-543,613). RESULTS: The contents of inflammasome proteins were gradually increased after cerebral ischemia/reperfusion (I/R). EA stimulus attenuated NLRP3 inflammasome mediated inflammatory reaction and regulated the balance between proinflammatory factors and anti-inflammatory cytokines. The agonist of α7nAChR induced similar neuroprotective effects as EA stimulus. In contrast, α7nAChR antagonist reversed not only the neuroprotective effects, but also the inhibitory effects of NLRP3 inflammasome and the regulatory effects on the balance between proinflammatory factors and anti-inflammatory cytokines. CONCLUSIONS: These results provided compelling evidence that α7nAChR played a pivotal role in regulating the activation and expression of NLRP3 inflammasome in neurons after cerebral I/R. These findings highlighted a novel anti-inflammatory mechanism of EA stimulus by α7nAChR modulating the inhibition of NLRP3 inflammasome, suggesting that α7nAChR-dependent cholinergic anti-inflammatory system and NLRP3 inflammasome in neurons might act as potential therapeutic targets in EA induced neuroprotection against cerebral ischemic injury.

Probenecid Relieves Cerebral Dysfunction of Sepsis by Inhibiting Pannexin 1-Dependent ATP Release.

Acute brain dysfunction and the following neurological manifestation are common complications in septic patients, which are associated with increased morbidity and mortality. However, the therapeutic strategy of this disorder remains a major challenge. Given the emerging role of a clinically approved drug, probenecid (PRB) has been recently identified as an inhibitor of pannexin 1 (PANX1) channel, which restrains extracellular ATP release-induced purinergic pathway activation and inflammatory response contributing to diverse pathological processes. In this study, we explored whether PRB administration attenuated neuroinflammatory response and cognitive impairment during sepsis. In mice suffered from cecal ligation and puncture (CLP)-induced sepsis, treatment with PRB improved memory retention and lessened behavioral deficits. This neuroprotective effect was coupled with restricted overproduction of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and interleukin (IL)-1ß in the hippocampus. Since this damped neuroinflammation was replicated by inhibition of ATP release, it suggested that PANX1 channel modulates a purinergic-related pathway contributing to the neurohistological damage. Therefore, we identified PRB could be a promising therapeutic approach for the therapy of cerebral dysfunction of sepsis.

Muscle-derived autologous mitochondrial transplantation: A novel strategy for treating cerebral ischemic injury.

The available evidence showed that mitochondrial transfer by releasing the extracellular vesicles containing mitochondria from astrocytes to neurons exerted a neuroprotective effect after stroke. Whether extracellular mitochondrial replenishment could rescue the tissues from cerebral ischemic injury still needs to be explored completely. It was hypothesized that the augmentation of mitochondrial damage after cerebral ischemia could be resolved by timely replenishment of exogenous mitochondria. A stroke model of middle cerebral artery occlusion (MCAO) was used in this study to verify this hypothesis. This study found that the number of extracellular mitochondria increased in rat cerebrospinal fluid after MCAO, and a higher proportion of mitochondria were associated with good neurological outcomes. Following 90-min ischemia, autologously derived mitochondria (isolated from autologous pectoralis major) or vehicle alone was infused directly into the lateral ventricles, and the rats were allowed to recover for 4 weeks. A plenty of infused mitochondria were found to be distributed in the boundary and ischemic penumbra areas. Furthermore, the transplantation of mitochondria reduced cellular oxidative stress and apoptosis, attenuated reactive astrogliosis, and promoted neurogenesis after stroke. Moreover, the transplantation of mitochondria decreased brain infarct volume and reversed neurological deficits. The findings suggested that the delivery of mitochondria through the lateral ventricles resulted in their widespread distribution throughout the brain and exerted a neuroprotective effect after ischemia-reperfusion injury.

MARCH5 RNA promotes autophagy, migration, and invasion of ovarian cancer cells.

MARCH5 is a crucial regulator of mitochondrial fission. However, the expression and function of MARCH5 in ovarian cancer have not been determined. This study investigated the expression and function of MARCH5 in ovarian cancer with respect to its potential role in the tumorigenesis of the disease as well as its usefulness as an early diagnostic marker. We found that the expression of MARCH5 was substantially upregulated in ovarian cancer tissue in comparison with the normal control. Silencing MARCH5 in SKOV3 cells decreased TGFB1-induced cell macroautophagy/autophagy, migration, and invasion in vitro and in vivo, whereas the ectopic expression of MARCH5 in A2780 cells had the opposite effect. Mechanistic investigations revealed that MARCH5 RNA may function as a competing endogenous RNA (ceRNA) to regulate the expression of SMAD2 and ATG5 by competing for MIR30A. Knocking down SMAD2 or ATG5 can block the effect of MARCH5 in A2780 cells. Also, silencing the expression of MARCH5 in SKOV3 cells can inhibit the TGFB1-SMAD2/3 pathway. In contrast, the ectopic expression of MARCH5 in A2780 cells can activate the TGFB1-SMAD2/3 pathway. In turn, the TGFB1-SMAD2/3 pathway can regulate MARCH5 and ATG5 through MIR30A. Overall, the results of this study identified MARCH5 as a candidate oncogene in ovarian cancer and a potential target for ovarian cancer therapy.