Enhanced Cardiomyocyte NLRP3 Inflammasome-Mediated Pyroptosis Promotes d-Galactose-Induced Cardiac Aging.

BACKGROUND: Cardiac aging represents an independent risk factor for aging-associated cardiovascular diseases. Although evidence suggests an association between NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome formation and numerous cardiovascular diseases, its role in cardiac aging remains largely unclear. METHODS AND RESULTS: The longevity of mice with wild-type and NLRP3 knockout (NLRP3-/-) genotypes was assessed, with or without d-galactose treatment. Cardiac function was evaluated using echocardiography, and cardiac histopathology was examined through hematoxylin and eosin and Masson's trichrome staining. Senescence-associated ß-galactosidase (SA-ß-gal) staining was employed to detect cardiac aging. Western blotting was used to assess aging-related proteins (p53, p21) and pyroptosis-related proteins. Additionally, dihydroethidium staining, lactate dehydrogenase release, and interleukin-1ß ELISA assays were performed, along with measurements of total superoxide dismutase and malondialdehyde levels. In vitro, H9c2 cells were exposed to d-galactose for 24 hours in the absence or presence of N-acetyl-l-cysteine (reactive oxygen species inhibitor), BAY-117082 (nuclear factor κ-light-chain enhancer of activated B cells inhibitor), MCC950 (NLRP3 inhibitor), and VX-765 (Caspase-1 inhibitor). Immunofluorescence staining was employed to detect p53, gasdermin D, and apoptosis-associated speck-like protein proteins. Intracellular reactive oxygen species levels were assessed using fluorescence microscopy and flow cytometry. Senescence-associated ß-galactosidase staining and Western blotting were also employed in vitro for the same purpose. The results showed that NLRP3 upregulation was implicated in aging and cardiovascular diseases. Inhibition of NLRP3 extended life span, mitigated the aging phenotype, improved cardiac function and blood pressure, ameliorated lipid metabolism abnormalities, inhibited pyroptosis in cardiomyocytes, and ultimately alleviated cardiac aging. In vitro, the inhibition of reactive oxygen species, nuclear factor κ-light-chain enhancer of activated B cells, NLRP3, or caspase-1 attenuated NLRP3 inflammasome-mediated pyroptosis. CONCLUSIONS: The reactive oxygen species/nuclear factor κ-light-chain enhancer of activated B cells/NLRP3 signaling pathway loop contributes to d-galactose-treated cardiomyocyte senescence and cardiac aging.

[Etiology composition and prognosis of pediatric chronic critical illness in a pediatric intensive care unit]. / å„¿ç«¥é‡ç—‡ç›‘æŠ¤å®¤å„¿ç«¥æ ¢æ€§å±é‡ç—‡çš„ç— å› ç»„æˆåŠé¢„åŽåˆ†æž.

OBJECTIVES: To explore the etiology composition and outcomes of pediatric chronic critical illness (PCCI) in the pediatric intensive care unit (PICU). METHODS: The children who were hospitalized in the PICU of Dongguan Children's Hospital Affiliated to Guangdong Medical University and met the diagnostic criteria for PCCI from January 2017 to December 2022 were included in the study. The etiology of the children was classified based on their medical records and discharge diagnoses. Relevant clinical data during hospitalization were collected and analyzed. RESULTS: Among the 3 955 hospitalized children in the PICU from January 2017 to December 2022, 321 cases (8.12%) met the diagnostic criteria for PCCI. Among the 321 cases, the most common etiology was infection (71.3%, 229 cases), followed by unintentional injury (12.8%, 41 cases), postoperation (5.9%, 19 cases), tumors/immune system diseases (5.0%, 16 cases), and genetic and chromosomal diseases (5.0%, 16 cases). Among the 321 cases, 249 cases (77.6%) were discharged after improvement, 37 cases (11.5%) were discharged at the request of the family, and 35 cases (10.9%) died in the hospital. Among the deaths, infection accounted for 74% (26/35), unintentional injury accounted for 17% (6/35), tumors/immune system diseases accounted for 6% (2/35), and genetic and chromosomal diseases accounted for 3% (1/35). From 2017 to 2022, the proportion of PCCI in PICU diseases showed an increasing trend year by year (P<0.05). Among the 321 children with PCCI, there were 148 infants and young children (46.1%), 57 preschool children (17.8%), 54 school-aged children (16.8%), and 62 adolescents (19.3%), with the highest proportion in the infant and young children group (P<0.05). The in-hospital mortality rates of the four age groups were 14.9% (22/148), 8.8% (5/57), 5.6% (3/54), and 8.1% (5/62), respectively. The infant and young children group had the highest mortality rate, but there was no statistically significant difference among the four groups (P>0.05). CONCLUSIONS: The proportion of PCCI in PICU diseases is increasing, and the main causes are infection and unintentional injury. The most common cause of death in children with PCCI is infection. The PCCI patient population is mainly infants and young children, and the in-hospital mortality rate of infant and young children with PCCI is relatively high.

Specific labeling and identification of bacteria based on concentration-dependent carbon dot staining combined with hyperspectral imaging.

Concentration-dependent carbon dot (CD) fluorescence was developed and utilized alongside hyperspectral microscopy as a specific labeling and identification technique for bacteria. Staining revealed that the CD concentration within cells depended on the characteristic intracellular environment of the species. Therefore, based on the concentration dependence of the CD fluorescence, different bacterial species were specifically labeled. Hyperspectral microscopy captured subtle fluorescence variations to identify bacteria. Method validation using Bacillus subtilis and Bacillus licheniformis succeeded with an identification accuracy of 99%. As a simple, rapid method for labeling and identifying bacterial species in mixtures, this technique has excellent potential for bacterial community studies.

Etiology composition and prognosis of pediatric chronic critical illness in a pediatric intensive care unit / 中国当代儿科杂志

OBJECTIVES@#To explore the etiology composition and outcomes of pediatric chronic critical illness (PCCI) in the pediatric intensive care unit (PICU).@*METHODS@#The children who were hospitalized in the PICU of Dongguan Children's Hospital Affiliated to Guangdong Medical University and met the diagnostic criteria for PCCI from January 2017 to December 2022 were included in the study. The etiology of the children was classified based on their medical records and discharge diagnoses. Relevant clinical data during hospitalization were collected and analyzed.@*RESULTS@#Among the 3 955 hospitalized children in the PICU from January 2017 to December 2022, 321 cases (8.12%) met the diagnostic criteria for PCCI. Among the 321 cases, the most common etiology was infection (71.3%, 229 cases), followed by unintentional injury (12.8%, 41 cases), postoperation (5.9%, 19 cases), tumors/immune system diseases (5.0%, 16 cases), and genetic and chromosomal diseases (5.0%, 16 cases). Among the 321 cases, 249 cases (77.6%) were discharged after improvement, 37 cases (11.5%) were discharged at the request of the family, and 35 cases (10.9%) died in the hospital. Among the deaths, infection accounted for 74% (26/35), unintentional injury accounted for 17% (6/35), tumors/immune system diseases accounted for 6% (2/35), and genetic and chromosomal diseases accounted for 3% (1/35). From 2017 to 2022, the proportion of PCCI in PICU diseases showed an increasing trend year by year (P<0.05). Among the 321 children with PCCI, there were 148 infants and young children (46.1%), 57 preschool children (17.8%), 54 school-aged children (16.8%), and 62 adolescents (19.3%), with the highest proportion in the infant and young children group (P<0.05). The in-hospital mortality rates of the four age groups were 14.9% (22/148), 8.8% (5/57), 5.6% (3/54), and 8.1% (5/62), respectively. The infant and young children group had the highest mortality rate, but there was no statistically significant difference among the four groups (P>0.05).@*CONCLUSIONS@#The proportion of PCCI in PICU diseases is increasing, and the main causes are infection and unintentional injury. The most common cause of death in children with PCCI is infection. The PCCI patient population is mainly infants and young children, and the in-hospital mortality rate of infant and young children with PCCI is relatively high.

D-galactose-induced cardiac ageing: A review of model establishment and potential interventions.

Cardiovascular disease (CVD) is highly prevalent in an ageing society. The increased incidence and mortality rates of CVD are global issues endangering human health. There is an urgent requirement for understanding the aetiology and pathogenesis of CVD and developing possible interventions for preventing CVD in ageing hearts. It is necessary to select appropriate models and treatment methods. The D-galactose-induced cardiac ageing model possesses the advantages of low mortality, short time and low cost and has been increasingly used in the study of cardiovascular diseases in recent years. Therefore, understanding the latest progress in D-galactose-induced cardiac ageing is valuable. This review highlights the recent progress and potential therapeutic interventions used in D-galactose-induced cardiac ageing in recent years by providing a comprehensive summary of D-galactose-induced cardiac ageing in vivo and in vitro. This review may serve as reference literature for future research on age-related heart diseases.

[Effect of sequential sedation and analgesia in preventing delirium and withdrawal symptoms in children after ventilator weaning]. / åºè´¯é•‡ç—›é•‡é™å¯¹é¢„é˜²è¡Œæœºæ¢°é€šæ°”æ²»ç–—æ‚£å„¿æ’¤æœºåŽè°µå¦„åŠæ’¤è¯ååº”çš„æ•ˆæžœç ”ç©¶.

OBJECTIVES: To investigate the effect of sequential sedative and analgesic drugs in preventing delirium and withdrawal symptoms in children after ventilator weaning. METHODS: A retrospective analysis was performed on 61 children who were admitted and received mechanical ventilation support for ≥5 days in the Pediatric Intensive Care Unit of Dongguan Children's Hospital Affiliated to Guangdong Medical University from December 2019 to September 2021. The children were divided into a control group (30 children with no maintenance of analgesic and sedative drugs after ventilator weaning) and an observation group (31 children with sequential sedative and analgesic drugs maintained for 48 hours after ventilator weaning). The two groups were compared in terms of the Sophia Observation Withdrawal Symptoms Scale (SOS) score, the Pediatric Delirium Scale (PD) score, the Richmond Agitation-Sedation Scale (RASS) score, and the incidence rates of delirium or withdrawal symptoms at 24 and 72 hours after ventilator weaning. RESULTS: There was no significant difference in the incidence rate of delirium at 24 hours and 72 hours after ventilator weaning between the two groups (P>0.05). Compared with the control group, the observation group had significantly lower incidence rate of withdrawal symptoms and scores of SOS, PD, and RASS scales at 24 hours and 72 hours after ventilator weaning (P<0.01). CONCLUSIONS: Sequential sedation and analgesia after ventilator weaning can reduce the incidence of withdrawal symptoms within 72 hours after ventilator weaning, but it cannot reduce the incidence rate of delirium.

[Application of ultrasound-guided central venous catheterization at various sites in infants with shock]. / è¶ å£°å¼•å¯¼ä¸‹ä¸åŒéƒ¨ä½ä¸­å¿ƒé™è„‰ç½®ç®¡åœ¨å©´å¹¼å„¿ä¼‘å ‹ä¸­çš„åº”ç”¨åŠä¸´åºŠåˆ†æž.

OBJECTIVES: To study the clinical characteristics of ultrasound-guided central venous catheterization at various sites in infants with shock, and to explore how to quickly select the site for central venous puncture in infants with shock. METHODS: The medical data of 112 infants who were diagnosed with shock and underwent central venous catheterization in the Pediatric Intensive Care Unit, Dongguan Children's Hospital Affiliated to Guangdong Medical University, from January 2016 to December 2020 were reviewed retrospectively. The patients were divided into an ultrasound group (n=70) and a body surface location group (n=42) according to whether the catheterization was carried out under ultrasound guidance. The application of ultrasound-guided catheterization at various sites in infants was summarized and analyzed, and the success rate of one-time puncture, overall success rate, catheterization time, and complications were compared between these sites. RESULTS: Compared with the body surface location group, the ultrasound group had a significantly higher success rate of one-time puncture, a significantly shorter catheterization time, and a significantly reduced incidence rate of complications in internal jugular vein and femoral vein catheterizations (P<0.05). In the ultrasound group, the proportion of internal jugular vein catheterization was the highest (51%, 36/70), followed by femoral vein catheterization (33%, 23/70), and subclavian vein catheterization (16%, 11/70). For the comparison between different puncture sites under ultrasound guidance, internal jugular vein catheterization showed the shortest time of a successful catheterization [5.5 (5.0, 6.5) minutes] (P<0.05). There was no significant difference in the incidence rate of complications among the different puncture sites groups (P>0.05). CONCLUSIONS: In infants with shock, ultrasound-guided internal jugular vein catheterization can be used as the preferred catheterization method for clinicians.

Identification of DAPI-stained normal, inflammatory, and carcinoma hepatic cells based on hyperspectral microscopy.

Gross chromatin imbalance and high DNA content are distinct features of various types of cancer cells. However, severe inflammation can also produce similar symptoms in cells. In this study, normal, inflammatory, and carcinoma hepatic cells were stained with 4',6-diamidino-2-phenylindole (DAPI) and investigated by hyperspectral microscopy. DAPI is a DNA-sensitive fluorochrome. Therefore, the differences in the cellular DNA of the samples can be revealed by the corresponding fluorescence. Our experimental results demonstrate that although chromosomal disorder and high DNA content both occur in severely inflammatory and carcinoma hepatic cells, there is still a slight difference in their DNA, making their fluorescent intensity and even their spectral shapes distinguishable. Based on these spectral features, we developed a method for the precise identification of normal, inflammatory, and carcinoma hepatic cells in the field of view. The identification accuracy for these three types of cells was 99.8%. We believe that examination that combines DAPI staining with hyperspectral microscopy is a potential method for the identification and investigation of various types of cancer tissues.

Effect of fluid load on the prognosis of children with sepsis-associated acute kidney injury undergoing continuous renal replacement therapy. / ä¸åŒæ¶²ä½“è´Ÿè·çŠ¶æ€å¯¹æŒç»­è‚¾æ›¿ä»£æ²»ç–—è„“æ¯’ç—‡ç›¸å ³æ€¥æ€§è‚¾æŸä¼¤æ‚£å„¿é¢„åŽçš„å½±å“.

OBJECTIVES: To evaluate the effect of fluid load on the prognosis of children with sepsis-associated acute kidney injury (AKI) undergoing continuous renal replacement therapy (CRRT). METHODS: A total of 121 children who underwent CRRT for sepsis-associated AKI from August 2018 to March 2021 were enrolled in the retrospective study. According to the fluid load from admission or disease progression to CRRT, they were divided into three groups: low fluid load (fluid load: <5%; n=35), high fluid load (fluid load: 5% - <10%; n=35), and fluid overload (fluid load: ≥10%; n=51). Baseline data and clinical biochemical data before CRRT were collected for comparison and analysis. The Kaplan-Meier survival curve analysis was used for comparison of 28-day survival between groups. The multivariate logistic regression model was used to identify the influencing factors for the prognosis of the children. RESULTS: The survival analysis showed that the fluid overload group had a significantly higher 28-day mortality rate than the low fluid load and high fluid load groups (P<0.05). The multivariate logistic regression analysis showed that an increase in fluid overload volume was a risk factor for increased 28-day mortality in the fluid overload group, while earlier initiation of CRRT was a protective factor (P<0.05). CONCLUSIONS: Fluid overload before CRRT may increase the mortality in children with sepsis-associated AKI, and CRRT should be performed for these children as early as possible.

Application of ultrasound-guided central venous catheterization at various sites in infants with shock / 中国当代儿科杂志

OBJECTIVES@#To study the clinical characteristics of ultrasound-guided central venous catheterization at various sites in infants with shock, and to explore how to quickly select the site for central venous puncture in infants with shock.@*METHODS@#The medical data of 112 infants who were diagnosed with shock and underwent central venous catheterization in the Pediatric Intensive Care Unit, Dongguan Children's Hospital Affiliated to Guangdong Medical University, from January 2016 to December 2020 were reviewed retrospectively. The patients were divided into an ultrasound group (n=70) and a body surface location group (n=42) according to whether the catheterization was carried out under ultrasound guidance. The application of ultrasound-guided catheterization at various sites in infants was summarized and analyzed, and the success rate of one-time puncture, overall success rate, catheterization time, and complications were compared between these sites.@*RESULTS@#Compared with the body surface location group, the ultrasound group had a significantly higher success rate of one-time puncture, a significantly shorter catheterization time, and a significantly reduced incidence rate of complications in internal jugular vein and femoral vein catheterizations (P<0.05). In the ultrasound group, the proportion of internal jugular vein catheterization was the highest (51%, 36/70), followed by femoral vein catheterization (33%, 23/70), and subclavian vein catheterization (16%, 11/70). For the comparison between different puncture sites under ultrasound guidance, internal jugular vein catheterization showed the shortest time of a successful catheterization [5.5 (5.0, 6.5) minutes] (P<0.05). There was no significant difference in the incidence rate of complications among the different puncture sites groups (P>0.05).@*CONCLUSIONS@#In infants with shock, ultrasound-guided internal jugular vein catheterization can be used as the preferred catheterization method for clinicians.

Effect of fluid load on the prognosis of children with sepsis-associated acute kidney injury undergoing continuous renal replacement therapy / 中国当代儿科杂志

OBJECTIVES@#To evaluate the effect of fluid load on the prognosis of children with sepsis-associated acute kidney injury (AKI) undergoing continuous renal replacement therapy (CRRT).@*METHODS@#A total of 121 children who underwent CRRT for sepsis-associated AKI from August 2018 to March 2021 were enrolled in the retrospective study. According to the fluid load from admission or disease progression to CRRT, they were divided into three groups: low fluid load (fluid load: <5%; n=35), high fluid load (fluid load: 5% - <10%; n=35), and fluid overload (fluid load: ≥10%; n=51). Baseline data and clinical biochemical data before CRRT were collected for comparison and analysis. The Kaplan-Meier survival curve analysis was used for comparison of 28-day survival between groups. The multivariate logistic regression model was used to identify the influencing factors for the prognosis of the children.@*RESULTS@#The survival analysis showed that the fluid overload group had a significantly higher 28-day mortality rate than the low fluid load and high fluid load groups (P<0.05). The multivariate logistic regression analysis showed that an increase in fluid overload volume was a risk factor for increased 28-day mortality in the fluid overload group, while earlier initiation of CRRT was a protective factor (P<0.05).@*CONCLUSIONS@#Fluid overload before CRRT may increase the mortality in children with sepsis-associated AKI, and CRRT should be performed for these children as early as possible.

Effect of sequential sedation and analgesia in preventing delirium and withdrawal symptoms in children after ventilator weaning / 中国当代儿科杂志

OBJECTIVES@#To investigate the effect of sequential sedative and analgesic drugs in preventing delirium and withdrawal symptoms in children after ventilator weaning.@*METHODS@#A retrospective analysis was performed on 61 children who were admitted and received mechanical ventilation support for ≥5 days in the Pediatric Intensive Care Unit of Dongguan Children's Hospital Affiliated to Guangdong Medical University from December 2019 to September 2021. The children were divided into a control group (30 children with no maintenance of analgesic and sedative drugs after ventilator weaning) and an observation group (31 children with sequential sedative and analgesic drugs maintained for 48 hours after ventilator weaning). The two groups were compared in terms of the Sophia Observation Withdrawal Symptoms Scale (SOS) score, the Pediatric Delirium Scale (PD) score, the Richmond Agitation-Sedation Scale (RASS) score, and the incidence rates of delirium or withdrawal symptoms at 24 and 72 hours after ventilator weaning.@*RESULTS@#There was no significant difference in the incidence rate of delirium at 24 hours and 72 hours after ventilator weaning between the two groups (P>0.05). Compared with the control group, the observation group had significantly lower incidence rate of withdrawal symptoms and scores of SOS, PD, and RASS scales at 24 hours and 72 hours after ventilator weaning (P<0.01).@*CONCLUSIONS@#Sequential sedation and analgesia after ventilator weaning can reduce the incidence of withdrawal symptoms within 72 hours after ventilator weaning, but it cannot reduce the incidence rate of delirium.

Classification of two species of Gram-positive bacteria through hyperspectral microscopy coupled with machine learning.

Gram stain is one of the most common techniques used to visualize bacteria under microscopy and classify bacteria into two large groups (Gram-positive and Gram-negative). However, such an inaccurate classification is unfavorable for bacterial research. For instance, soil-rhizosphere bacteria, Bacillus megaterium (B. megaterium) and Bacillus cereus (B. cereus) have different effects on plants, nonetheless, they are both Gram-positive and difficult to be differentiated. Here, we present a method to precisely classify Gram-positive bacteria via hyperspectral microscopy. The pH-value differences in the intracellular environment of various types of bacteria can lead to different ionization of the auxochrome of crystal violet (CV) molecules during the Gram stain process. Consequently, there is a subtle difference in the absorption peak of Gram-stained bacteria. Harnessing hyperspectral microscopy can capture this subtle difference and enable precise classification. Besides the spectral features, the spatial features were also used to improve the quality of bacterial identification. The results show that the classification accuracy of two species of Gram-positive bacteria, B. megaterium and B. cereus, is up to 98.06%. We believe this method can be used for other Gram-positive bacteria and Gram-negative bacteria, realizing a more elaborate classification for Gram-stained bacteria.

[Cloning and expression of triosephosphate isomerase gene of Boophilus microplus].

Total RNA was extracted from adult Boophilus microplus. RT-PCR was used to amplify the gene and the fragment was subcloned into the expression vector pET-28a. The cloned gene was expressed in E. coli Rosseta (DE3), induced by IPTG, and identified by SDS-PAGE. The results showed that the triosephosphate isomerase (tim) gene of B. microplus has 750bp and encodes 249 amino acids (GenBank No. JX112888). The cloned tim gene shares 99% homology with that in tick embryos. The relative molecular weight (M(r)) of the expressed recombinant protein is about 27 000.