Venomous snakebites in children: a 10 year experience in South China.

INTRODUCTION: Many studies have focused on snakebites in adults, but very few have described snakebites in children. METHODS: We reviewed the clinical characteristics and outcomes of children with venomous snakebites aged less than 15 years who presented to a regional medical centre in South China from January 2013 to December 2022. RESULTS: A total of 69 envenomed patients were analyzed in our study; 42 (60.9 per cent) patients were male, and 59 (85.5 per cent) reported lower limb bites. Most bites (89.8 per cent) occurred between April and October. Twenty-seven patients received first aid management, and 47 required admission to the general ward. Antivenom was administered to 58 patients, glucocorticoids to 43 patients, antibiotics to 48 patients, and tetanus antitoxin to 40 patients. No fatalities were reported. The most common snake identified was Trimeresurus albolabris. Four were classified as dry bites, 15 as mild, 43 as moderate, and seven as severe. The most common local signs were pain and swelling, while the most common systemic effects were haematological complications. Patients with high severity scores had significantly higher lactate dehydrogenase activities, creatine kinase isoenzyme activities, aspartate aminotransferase activities, D-dimer concentrations, prothrombin times and lower fibrinogen concentrations. In a receiver operating characteristic curve analysis of the values with the highest Youden index, the following cut-offs proved significant: lactate dehydrogenase activity > 248.1 U/L, creatine kinase isoenzyme activities > 17.5 U/L, fibrinogen concentration < 1,455 mg/L, D-dimer concentration > 437.0 µg/L, aspartate aminotransferase activity > 26.1 U/L, and prothrombin time > 15.2 seconds. DISCUSSION: This study provides insight into the epidemiology, clinical profile, and management of snakebites in children. Data from the present study were compared with those from our previous adult study. Limitations include that 50.7 per cent of our snakebites were attributed to Trimeresurus albolabris. Therefore, the results of our study may not be generalizable to all snakebites. CONCLUSION: The clinical symptoms were more severe in children than in adults in our previous study. Even though there were no fatalities, close monitoring should be performed to detect haematological and other potentially fatal complications promptly.

Proteomic characteristics of six snake venoms from the Viperidae and Elapidae families in China and their relation to local tissue necrosis.

Patients envenomed by snakes from the Viperidae and Elapidae families in China often have varying degrees of local tissue necrosis. Due to the relative clinical characteristics of local tissue necrosis and ulceration following envenoming, this study has analyzed the proteome of six snake venoms from the Viperidae and Elapidae family, and the toxin profiles of each snake were compared and correlated with the clinical manifestations that follow cytotoxic envenoming. Deinagkistrodon acutus and Naja atra envenomation induce severe ulceration, which is absent in Bungarus multicinctus envenomation and mild in the other three vipers. It is interesting to note that the proportion of c-type lectins (CTL) (20.63%) in Deinagkistrodon acutus venom was relatively high, which differs from the venom of other vipers. In addition, three-fingered toxin (3FTx) (2.15%) is present in the venom of Deinagkistrodon acutus, but has not been detected in the remaining three vipers. Snake venom metalloprotease (SVMP) (34.4%-44.7%), phospholipase A2 (PLA2) (9.81%-40.83%), and snake venom serine protease (SVSP) (9.44%-16.2%) represent the most abundant families of toxin in Viperidae venom. The Elapidae venom proteome was mainly composed of neurotoxins and cytotoxins, including 3FTx (39.28%-60.08%) and PLA2 (8.24%-58.95%) toxins, however, the proportion of CRISPS (26.36%) in Naja atra venom was relatively higher compared to Bungarus multicinctus venom. Significant differences in SVMP, SVSP, and 3FTx expression levels exist between the Viperidae and the Elapidae family. The main toxins responsible for the development of tissue necrosis and ulcerations following Viperidae envenoming are hematotoxins (SVSMP, SVSP) and myotoxins (PLA2). Deinagkistrodon acutus venom contains high levels of CTL and traces of 3FTx, leading to more severe local necrosis. However, Naja atra venom can also cause severe local necrosis through the effects of myotoxin (3FTx, CRISP, PLA2). Bungarus multicinctus venom does not contain myotoxins, resulting in pure systemic neurological manifestations no obvious necrosis of local tissue in patients.

Inhibition of ferroptosis alleviates chronic unpredictable mild stress-induced depression in mice via tsRNA-3029b.

Depression is a common mental illness. Ferroptosis is a form of cell death that may be responsible for neurological disease, but the role of ferroptosis in depression remains unclear. tRNA-derived small RNA (tsRNA) is an emerging non-coding small RNA, making it an important medium for studying neurological diseases. Chronic unpredictable mild stress (CUMS) was used to construct the depression model in mice, which was treated with ferrostatin-1 (Fer-1). Classical behavioral test, immunofluorescence and small RNA sequencing were used to detect depression-like behaviors, neuronal proliferation and the expression profile of tsRNAs in mice, respectively. The primary neuronal cell damage model was constructed by corticosterone (CORT), and the function of key tsRNA was investigated by quantitative real-time PCR, western blot and CCK-8 assays. Here, Fer-1 reduced the depression-like behavior of CUMS-induced mice and promoted neuronal growth. In addition, CUMS caused the disorder of tsRNA expression profile in hippocampal tissues of mice, and Fer-1 alleviated the abnormal tsRNA expression, among which tsRNA-3029b was an effective target. In vitro experiments manifested that ROS accumulation and decreased expression of SLC7A11 and GPX4 were found in CORT-induced depression-like cell model, suggesting that ferroptosis was involved in neuronal injury. However, inhibition of tsRNA-3029b suppressed neuronal cell ferroptosis and facilitated neuronal regeneration. In conclusion, Fer-1 showed an antidepressant effect in CUMS-induced mice and alleviated the abnormal expression profile of tsRNA. tsRNA-3029b was a key target in depression, and silencing of tsRNA-3029b reduced the occurrence of ferroptosis and protected neurons from injury, which may provide novel target for the treatment of depression.

The Arabidopsis thaliana integrin-like gene AT14A improves drought tolerance in Solanum lycopersicum.

Using effective genes to improve crop stress tolerance through genetic engineering is an important way to stabilize crop yield and quality across complex climatic environments. Integrin-like AT14A, as a continuum of the cell wall-plasma membrane-cytoskeleton, functions in the regulation of cell wall synthesis, signal transduction, and the response to stress. In this study, AT14A was overexpressed in Solanum lycopersicum L. In transgenic plants, both chlorophyll content and net photosynthetic rate increased. Physiological experiments suggested that the proline content and antioxidant enzyme (superoxide dismutase, catalase, peroxidase) activities of the transgenic line were significantly higher than those of wild-type plants under stress, which contributed to the enhanced water retention capacity and free radical scavenging ability of the transgenic line. Transcriptome analysis revealed that AT14A enhanced drought tolerance by regulating waxy cuticle synthesis genes, such as 3-ketoacyl-CoA synthase 20 (KCS20), non-specific lipid-transfer protein 2 (LTP2), antioxidant enzyme system genes peroxidase 42-like (PER42), and dehydroascorbate reductase (DHAR2). AT14A regulates expression of Protein phosphatase 2 C 51 (PP2C 51) and ABSCISIC ACID-INSENSITIVE 5 (ABI5) to participate in ABA pathways to enhance drought tolerance. In conclusion, AT14A effectively improved photosynthesis and enhanced drought tolerance in S. lycopersicum.

Saussurea involucrata PIP2;4 improves growth and drought tolerance in Nicotiana tabacum by increasing stomatal density and sensitivity.

Aquaporins, the major facilitators of water transport across membranes, are involved in growth and development and adaptation to drought stress in plants. In this study, a plasma membrane intrinsic protein (SiPIP2;4) was cloned from Saussurea involucrata, a cold-tolerant hardy herb. The expression of SiPIP2;4 increased the stomatal density and sensitivity of tobacco (Nicotiana tabacum), thus, affecting the plant's growth and resistance to the diverse water environment. The higher stomatal density under well-watered conditions effectively promoted the photosynthetic rate, which led to the rapid growth of transgenic lines. The stomata in the transgenic lines responded more sensitively to the vapor pressure deficit than the wild-type under different levels of ambient humidity. Their stomatal apertures positively correlated with the ambient humidity. Under drought conditions, the overexpression of SiPIP2;4 promoted rapid stomatal closure, reduced water dissipation, and enhanced drought tolerance. These results indicate that SiPIP2;4 regulates the density and sensitivity of plant stomata, thus, playing an important role in balancing plant growth and stress tolerance. This suggests that SiPIP2;4 has the potential to serve as a genetic resource for crop improvement.

Adverse reactions to four types of monovalent antivenom used in the treatment of snakebite envenoming in South China.

To evaluate the adverse reactions associated with the four types of monovalent antivenom currently used in China, we retrospectively analysed the data of all patients admitted for snakebites who received antivenom treatment at the main institution for the treatment of venomous snakebites in Guangzhou from January 2013 to December 2021. A total of 1658 patients were analysed in our study, and 60.7% (n = 1007) of the snakebite patients received antivenom treatment. The incidence rate of adverse reactions that occurred after the administration of all types of monovalent antivenom was 4.9% (n = 49), and the incidence rate of acute adverse reactions was 2.7% (n = 27). The number of adverse reactions that occurred was 38/744 (5.1%) in patients who received prophylactic application of glucocorticoids alone and 10/217 (4.6%) in those who received a combination of antihistamines and glucocorticoids (P = 0.83). The average doses of the antivenoms used in patients exhibiting acute adverse reactions and serum sickness were 3.31 ± 0.75 vials and 2.36 ± 0.26 vials, respectively (P = 0.28). The antivenom skin test showed high specificity (98.3%, 95% CI: 97.24%-99.01%) but low sensitivity (14.3%, 95% CI: 6.41%-27.86%). Our results showed that the four types of monovalent antivenom were safe. No significant difference was observed between the use of glucocorticoids alone and the use of antihistamines combined with glucocorticoids as premedication for the prevention of adverse reactions. Reducing the dose of antivenoms or reducing the combination of antivenoms did not help to reduce the occurrence of adverse reactions. Skin testing should not be recommended due to its low sensitivity.

[Effect of Competitive Antagonist of Transmembrane Glutamine Flux V-9302 on Apoptosis of Acute Myeloid Leukemia Cell Lines HL-60 and KG-1].

OBJECTIVE: To investigate the effect of V-9302 (an antagonist of transmembrane glutamine flux) on the proliferation and apoptosis of acute myeloid leukemia cells HL-60 and KG-1. METHODS: HL-60 and KG-1 cells at logarithmic phase were treated by different concentrations of V-9302. CCK-8 assay was used to detect the proliferation of the cells. Annexin V-FITC / PI double staining flow cytometry was used to detect the apoptosis of HL-60 and KG-1 cells. The expressions of BAX, BCL-2 and Caspase3 were detected by RT-qPCR and Western blot. RESULTS: V-9302 could significantly inhibit the growth of HL-60 and KG-1 cells. The concentration of V-9302 at 10, 20 µmol/L could significantly promote the apoptosis of HL-60 and KG-1 cells(P<0.05). The results of apoptosis related gene detection showed that when V-9302 was applied to HL-60 and KG-1 cell lines at 10 and 20 µmol/L, the expression levels of Pro-apoptotic protein genes BAX and Caspase3 in HL-60 and KG-1 were significantly higher than those in control group (P<0.05), while the expression level of anti-apoptotic protein gene BCL-2 was significantly lower than that in the control group (P<0.05). The results of Western blot were basically consistent with that of RT-qPCR. CONCLUSION: Competitive antagonist of transmembrane glutamine flux V-9302 can significantly promote the apoptosis of acute myeloid leukemia cell lines HL-60 and KG-1.

Alterations of long non-coding RNA and mRNA profiles associated with extracellular matrix homeostasis and vascular aging in rats.

Vascular aging has been closely associated with various cardiovascular disorders; however, its molecular mechanism remains poorly understood. In our study, RNA sequencing was utilized to explore the expression profiles of long non-coding RNAs (lncRNAs) and mRNAs in the thoracic aortas of young (3 weeks) and old (16 weeks) rats. Functional categorization of differentially expressed mRNAs was evaluated using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases, and lncRNA-microRNA-mRNA networks was constructed using Cytoscape software. In addition, three upregulated and three downregulated lncRNAs were further confirmed by quantitative reverse transcriptase-polymerase chain reaction. A total of 36 lncRNAs and 922 mRNAs were differential expression in the thoracic aortas of young and older rats. In addition, we found differentially expressed mRNAs that were enriched in multiple biological processes and signaling pathways associated with angiogenesis, such as extracellular matrix-receptor interaction and adenosine 3',5'-monophosphate-activated protein kinase (AMPK) signaling. Moreover, AABR07013558.1, AABR07014823.1, and AABR07031489.1 were upregulated and ABR07053849.3, AABR07067310.2, and AC111292.1 were downregulated in the thoracic aortas of older rats compared with the young ones. Therefore, our findings provide several potential lncRNAs and mRNAs and signaling pathways related to vascular aging, which provide new clue for underlying the improvement of vascular aging.

The diagnostic accuracy of procalcitonin in infectious patients after cardiac surgery: a systematic review and meta-analysis.

Cardiac surgery with cardiopulmonary bypass (CPB) induces an acute inflammatory response that may lead to a systemic inflammatory response syndrome. The interest in procalcitonin (PCT) in the diagnosis of bacterial infection in patients after cardiac surgery remains less defined. The aim of this meta-analysis is to prospectively examine the discriminatory power of PCT as markers of infection in hospitalized patients with after cardiac surgery. The bivariate generalized nonlinear mixed-effect model and the hierarchical summary receiver operating characteristic model were used to estimate the pooled sensitivity, specificity and summary receiver operating characteristic curve. The pooled sensitivity and specificity were 0.81 (95% CI 0.75-0.87) and 0.78 (95% CI 0.73-0.83), respectively. The pooled positive likelihood ratio, and negative likelihood ratio of PCT were 3.74 (95% CI 2.98-4.69) and 0.24 (95% CI 0.17-0.32), respectively. The pooled area under the summary receiver operating characteristic curve of PCT using the HSROC method was 0.87 (95% CI 0.84- 0.90). This study indicated that PCT is a promising marker for the diagnosis of sepsis for those patients who undergo cardiac surgery.

ZJ01, a Small Molecule Inhibitor of the Kelch-Like ECH-Associated Protein 1-Nuclear Factor Erythroid 2-Related Factor 2 (Keap1-Nrf2) Protein-Protein Interaction, Reduces Hyperoxic Acute Lung Injury in a Mouse Model.

BACKGROUND Hyperoxic acute lung injury (ALI) is a complication of ventilation in patients with respiratory failure. Nuclear factor erythroid-2-related factor 2 (Nrf2) has an important role in ALI. Kelch-like ECH-associated protein 1 (Keap1) binds to Nrf2. ZJ01 is a small molecule inhibitor of Keap1-Nrf2 protein-protein interaction (PPI) that can reduce Keap1-induced inhibition of Nrf2. This study aimed to investigate the effects of ZJ01 and the heme oxygenase-1 (HO-1) inhibitor, zinc protoporphyrin IX (ZnPP IX), in a mouse model of hyperoxic ALI. MATERIAL AND METHODS C57BL/6J mice included five study groups: the room air+vehicle-treated group; the room air+ZJ01 group; the hyperoxia+vehicle-treated group; the hyperoxia+ZJ01 group; and the hyperoxia+ZJ01+ZnPP IX group. ZJ01, ZnPP IX, or vehicle were given 1 h after the hyperoxia challenge. The lungs from the mice were harvested at 72 h following the hyperoxia challenge. RESULTS Hyperoxia exposure for 72 h increased the activity of myeloperoxidase, the lung water content, the levels of tumor necrosis factor-alpha (TNF-alpha), and matrix metalloprotease-9 (MMP-9) in the vehicle-treated mice. ZJ01 treatment reduced hyperoxia-induced inflammation and increased the activation of Nrf2 and HO-1 compared with the vehicle-treated mice. Histology of the lungs showed that ZJ01 treatment reduced the changes of hyperoxia-induced ALI. Pretreatment with ZnPP IX reversed the beneficial effect of ZJ01. CONCLUSIONS ZJ01, a Keap1-Nrf2 PPI inhibitor, reduced hyperoxic ALI in a mouse model through the Nrf2/HO-1 pathway.

Lymphotoxin beta receptor is associated with regulation of microRNAs expression and nuclear factor-kappa B activation in lipopolysaccharides (LPS)-stimulated vascular smooth muscle cells.

BACKGROUND: The aim of present study is to investigate the role of lymphotoxin beta receptor (Ltßr) in lipopolysaccharides (LPS)-induced inflammation in vascular smooth muscle cells (VSMCs) and whether its effects are mediated by modulating microRNAs (miRNAs) and nuclear factor-kappa B (NF-κB). METHODS: Mouse aortic smooth muscle cell (SMC) line (MOVAS cells) were transduced with short hairpin Ltßr (shLtßr) and mRNA and protein expression level of Ltßr were measured by qPCR and Western blot in shLtßr-transduced cells. Lentiviral vector-transduced (control) and lentiviral vector/shLtßr-transduced MOVAS cells were stimulated with LPS (1 µg/mL) for 0, 16, or 24 h. Then the mRNA and protein levels of Ltßr, interleukin-18 (IL-18), p-p65, p65 and vascular cell adhesion molecule 1 (VCAM-1) were measured by real-time quantitative polymerase chain reaction (qPCR), Western blot and enzyme-linked immunosorbent assay (ELISA). Different miRNAs expression in LPS-stimulated normal and shLtßr-transduced cells were detected by small RNA sequencing (smRNA-seq). RESULTS: The mRNA and protein expression of Ltßr was significantly downregulated in shLtßr-transduced cells. LPS-increased the mRNA and protein levels of Ltßr, IL-18, p-p65 and VCAM-1 in were attenuated by shLtßr transducing compared with LPS-stimulated control group. Moreover, LPS treatment induced 10 upregulated and 64 downregulated miRNAs in shLtßr-transduced cells compared with control cells. Moreover, miR-146b-5p and miR-27a-5p levels were significantly decreased in shLtßr-transduced cells. CONCLUSIONS: Our results show for the first time that the role of Ltßr in regulating inflammatory response in LPS-stimulated VSMCs via modulating miRNAs and NF-κB pathway. Our findings might provide valuable information with respect to better understanding in the treatment of cardiovascular diseases, such as atherosclerosis.

Interleukin 37 Suppresses M1 Macrophage Polarization Through Inhibition of the Notch1 and Nuclear Factor Kappa B Pathways.

Macrophage-orchestrated chronic inflammation plays an important role in cardiovascular disease, including accelerating the development of calcific aortic valve disease (CAVD). M1 and M2 macrophage polarization imbalances can alter intensity of inflammatory responses. Recombinant human interleukin 37 (IL-37) could be involved in regulating immune cell function to attenuate inflammation. This study aimed to identify IL-37 specifically modulates M1 polarization and investigate the underlying mechanism. Compared with normal valves, there are more M1 macrophages accumulation and less IL-37 expression in calcific aortic valves, which may indicate a negative relationship between IL-37 and M1 polarization. THP-1 cells could differentiate into resting macrophages with phorbol-12-myristate-13-acetate (PMA) and then polarize into M1 macrophages following treatment with lipopolysaccharide (LPS) and interferon gamma (IFN-γ). In vitro, recombinant human IL-37 attenuated the expression of inducible nitric oxide synthase (iNOS), CD11c, IL-6 and monocyte chemoattractant protein 1 (MCP-1) in M1 but augmented the expression of CD206 and IL-10 in M2. The suppression of M1 polarization was associated with the inhibition of the activation of the nuclear factor kappa B (NF-κB) and Notch1 signaling pathways. These results demonstrated that IL-37 inhibits the macrophages polarizing into M1 type via the inhibition of the Notch1 and nuclear factor kappa B pathways. In summary, IL-37 could be a potential therapeutic candidate for progressive CAVD by modulating M1 polarization and its orchestrated inflammation.