Inflammasome is a large protein complex activated upon cellular stress or microbial infection, which triggers maturation of pro-inflammatory cytokines interleukin-1β and interleukin-18 through caspase-1 activation. Nod-like receptor family protein 3 (NLRP3) is the most characterized inflammasome activated by various stimuli. However, the mechanism of its activation is unclear and its exact cellular localization is still unknown. We examined the potential co-localization of NLRP3 inflammasome with mitochondria and seven other organelles under adenosine triphosphate, nigericin or monosodium urate stimulation in mouse peritoneal macrophages using confocal microscopy approach. Our results revealed that the activated endogenous apoptosis-associated speck-like protein containing a CARD (ASC) pyroptosome forms in the cytoplasm and co-localizes with NLRP3 and caspase-1, but not with any of the organelles screened. This study indicates that the ASC pyroptosome universally localizes within the cytoplasm rather than with any specific organelles.
Adenosine Triphosphate ; pharmacology ; Animals ; Apoptosis Regulatory Proteins ; CARD Signaling Adaptor Proteins ; Carrier Proteins ; analysis ; metabolism ; Caspase 1 ; analysis ; metabolism ; Cytoplasm ; metabolism ; Cytoskeletal Proteins ; analysis ; metabolism ; Inflammasomes ; analysis ; metabolism ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Microscopy, Confocal ; Mitochondria ; metabolism ; NLR Family, Pyrin Domain-Containing 3 Protein ; Nigericin ; pharmacology ; Uric Acid ; pharmacology

Objective:To search and evaluate the literatures on the application of mechanical insufflation-exsufflation technique (MI-E) in patients with invasive mechanical ventilation in China and abroad, and to summarize the best evidence to provide evidence-based basis for clinical practice.Methods:The literatures related to the use of MI-E technique in invasive mechanical ventilation patients were searched from the establishment of the database to April 1, 2022 in BMJ Best Practice, UpToDate Clinical Advisor, Scottish Intercollegiate Guidelines Network (SIGN), Guidelines International Network (GIN), National Institute for Health and Care Excellence (NICE), National Guideline Clearinghouse (NGC), Registered Nurses' Association of Ontario (RNAO), medlive, Cochrane Library, Joana Briggs Instiute, Web of Science, PubMed, Embase, Cumulative Index to Nursing and Allied Health Literature (CINAHL), SinoMed, China National Knowledge Infrastructure (CNKI), Wanfang Database, China Science and Technology Journal Database, and Website of American Association for Respiratory Care, including guideline, expert consensus, clinical decision, evidence summary, systematic review and randomized controlled trial. The JBI Center for Evidence-Based Health Care Expert Consensus Evaluation Criteria (2016) was used to evaluate the quality of the included systematic reviews; the JBI Center for Evidence-Based Health Care Evaluation Criteria for Randomized Controlled Trials was used to evaluate the quality of the included randomized controlled trial. Two researchers independently evaluated the quality of literature, and extracted and summarized the evidence based on professional judgment.Results:A total of 7 literatures were enrolled, including 3 systematic reviews and 4 randomized controlled trials. After quality evaluation, 7 articles were all enrolled. Thirteen best evidences were formed from four aspects of indications, contraindications, parameter settings, and attention.Conclusions:The study summarizes the best evidence for the application of MI-E technique in invasive mechanical ventilation patients. It is recommended that medical staff undergo professional training, combined with their professional judgment as well as the patient's clinical specific conditions and willingness, and accurately apply MI-E technology to invasive mechanical ventilation patients.

Objective: To establish comprehensive laboratory reference intervals for Chinese children. Methods: This was a cross-sectional multicenter study. From June 2013 to December 2014, eligible healthy children aged from 6-month to 17-year were enrolled from 20 medical centers with informed consent. They were assessed by physical examination, questionnaire survey and abdominal ultrasound for eligibility. Fasting blood samples were collected and delivered to central laboratory. Measurements of 15 clinical laboratory parameters were performed, including estradiol (E2), testosterone(T), luteinizing hormone(LH), follicle-stimulating hormone(FSH), alanine transaminase(ALT), serum creatinine(Scr), cystatin C, immunoglobulin A(IgA), immunoglobulin G(IgG), immunoglobulin M(IgM), complement (C3, C4), alkaline phosphatase(ALP), uric acid(UA) and creatine kinase(CK). Reference intervals were established according to central 95% confidence intervals for reference population, stratified by age and sex. Results: In total, 2 259 children were enrolled. Finally, 1 648 children were eligible for this study, including 830 boys and 818 girls, at a mean age of 7.4 years. Age- and sex- specific reference intervals have been established for the parameters. Reference intervals of sex hormones increased gradually with age. Concentrations of ALT, cystatin C, ALP and CK were higher in children under 2 years old. Serum levels of sex hormones, creatinine, immunoglobin, CK, ALP and urea increased rapidly in adolescence, with significant sex difference. In addition, reference intervals were variable depending on assay methods. Concentrations of ALT detected by reagents with pyridoxal 5'-phosphate(PLP) were higher than those detected by reagents without PLP. Compared with enzymatic method, Jaffe assay always got higher results of serum creatinine, especially in children younger than 9 years old. Conclusion This study established age- and sex- specific reference intervals, for 15 clinical laboratory parameters based on defined healthy children.

Objective:To investigate the effect of early external diaphragm pacing on the diaphragm function and prognosis of patients with mechanical ventilation.Methods:A total of 47 patients receiving invasive mechanical ventilation in the Emergency Intensive Care Unit of Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University) from October 2019 to July 2021 were selected and randomly divided into the treatment group ( n=23) and control group ( n=24). The patients in the control group received routine clinical treatment. On this basis, the treatment group received external diaphragm pacing treatment every day from the third day of mechanical ventilation until weaning, and was followed up to 30 days after discharge. The diaphragm thickness, diaphragm excursion, diaphragm thickening fraction, mechanical ventilation time, number of weaning failures, length of hospital stay and death toll were compared between the two groups. Results:Compared with the control group, the diaphragm thickness [(0.21±0.05) cm vs. (0.16±0.05) cm], diaphragm excursion [(1.38±0.37) cm vs. (1.11±0.48) cm], and diaphragm thickening fraction [26% (19%, 32%) vs. 18.5% (10.25%, 20%)] in the treatment group increased significantly (all P<0.05). The mechanical ventilation time was shorter in the treatment group [10 (7, 15) d vs. 13 (10.25, 19) d], and the difference was statistically significant ( P<0.05). There were no significant differences in the number of weaning failures (7 vs. 9), length of hospital stay [22 (15 , 30) d vs.. 24 (17.25, 34.25) d] and deaths (8 vs. 8) (all P>0.05). Conclusions:Early application of external diaphragm pacing can improve the diaphragm function of patients with mechanical ventilation, delay the decline in diaphragm function, increase diaphragm excursion and diaphragm thickening fraction, and shorten the mechanical ventilation time.