Anti-inflammatory effects of paeoniflorin caused by regulation of the hif1a/miR-210/caspase1/GSDMD signaling pathway in astrocytes: a novel strategy for hypoxia-induced brain injury in rats.

Context: Hypoxia-induced injury is a classic symptom of obstructive sleep apnea hypopnea syndrome (OSAHS), which is a risk factor of various diseases, such as hypertension, heart failure and stroke. However, there is no effective therapy for hypoxia-induced injury or OSAHS due to the elusive mechanism involved.Objective: This study aimed to assess the effects of paeoniflorin on hypoxia-induced injury and explore the underlying mechanism.Materials and methods: Hypoxic models of SD rats and CTX-TNA2 cells were used to assess the effect of paeoniflorin, and the expressions of hif1a, miR-210, caspase1 and GSDMD were detected using western blots and RT-PCR. Plasmid transfection was performed to explore the role of miR-210 in the effect of paeoniflorin.Results: Firstly, we confirmed that hypoxia induced severe neuronal injury and an enhancement of inflammation in the rat brain, with elevated expression of caspase1, IL1b and IL18. In addition, the results showed an activation of astrocytes and an increased level of pyroptosis under hypoxic conditions, which suggested a critical role of pyroptosis in hypoxiainduced injury of the brain. Furthermore, we found that compared with the controls, paeoniflorin treatment improved hypoxia-induced pyroptosis in astrocytes. Moreover, we detected the activation of hif1a/miR-210 signaling in the effects of paeoniflorin on astrocytes. As expected, the expression of hif1a and miR-210 was significantly upregulated in astrocytes when exposed to hypoxia, while paeoniflorin treatment reversed these enhancements. After transfection of miR-210 mimics, the attenuation of pyroptosis induced by paeoniflorin was suppressed, which was accompanied by an increase of ROS levels, as well as LDH release, indicating a critical role of miR-210 in pyroptosis in astrocytes.Conclusions: Our findings demonstrated that paeoniflorin improved hypoxia-induced pyroptosis in astrocytes via depressing hif1a/miR-210/caspase1/GSDMD signaling, providing robust evidence for the treatment of hypoxic injury and OSAHS.HighlightsHypoxia induces severe injury and inflammatory response in the rat brain;Hypoxia enhanced pyroptotic level and led to an activation of astrocytes.;Paeoniflorin alleviates hypoxia-induced pyroptosis in astrocytes;Transfection of miR-210 mimics suppressed the effects of paeoniflorin on hypoxia-induced pyroptosis in astrocytes.

Chemerin-9 Peptide Enhances Memory and Ameliorates Aß1-42-Induced Object Memory Impairment in Mice.

Accumulating evidence suggests that the inhibition of neuroinflammation is a potential target for therapeutic or preventive strategies for Alzheimer's disease (AD). Chemerin has attracted particular attention for its role in the regulation of inflammation. In addition, amyloid ß1-42 (Aß1-42) can interact with chemokine-like receptor 1 (CMKLR1), the receptor for chemerin, and induce microglial chemotaxis. Meanwhile, CMKLR1 is expressed in the brain, and both chemerin and Aß1-42 share the same receptor. Thus, we hypothesized that chemerin (C9), a chemerin-derived nonapeptide, may have the potential to ameliorate Aß1-42 mediated AD disease progression. The results showed that an intracerebroventricular (i.c.v.) injection of C9 (8 µg/kg) facilitated memory formation and improved memory retention, as evidenced by the results of both the novel object recognition test (NOR) and object location recognition (OLR) tasks. These memory-enhancing effects of C9 were also observed after C9 (2 µg/kg) was infused into the hippocampus. Moreover, we found that treatment with C9 reversed the deficits in memory and learning ability induced by oligomeric Aß1-42. Meanwhile, C9 also significantly inhibited Aß1-42-induced increases in the levels of pro-inflammatory cytokines such as interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the hippocampus. The same results were obtained for Western blotting and enzyme-linked immunosorbent assay (ELISA) experiments. Finally, we observed that C9 did not affect locomotor activity, suggesting that its improvement of memory is not a false positive induced by hypolocomotion. In conclusion, C9 may facilitate memory formation, prolong memory retention, and ameliorate Aß1-42-induced memory impairment, suggesting that C9 may potentially represent a novel strategy for the treatment of AD.

Strain-level multidrug-resistant pathogenic bacteria in urban wastewater treatment plants: Transmission, source tracking and evolution.

Wastewater treatment plants (WWTPs) serve as reservoirs for various pathogens and play a pivotal role in safeguarding environmental safety and public health by mitigating pathogen release. Pathogenic bacteria, known for their potential to cause fatal infections, present a significant and emerging threat to global health and remain poorly understood regarding their origins and transmission in the environment. Using metagenomic approaches, we identified a total of 299 pathogens from three full-scale WWTPs. We comprehensively elucidated the occurrence, dissemination, and source tracking of the pathogens across the WWTPs, addressing deficiencies in traditional detection strategies. While indicator pathogens in current wastewater treatment systems such as Escherichia coli are effectively removed, specific drug-resistant pathogens, including Pseudomonas aeruginosa, Pseudomonas putida, and Aeromonas caviae, persist throughout the treatment process, challenging complete eradication efforts. The anoxic section plays a predominant role in controlling abundance but significantly contributes to downstream pathogen diversity. Additionally, evolution throughout the treatment process enhances pathogen diversity, except for upstream transmission, such as A. caviae str. WP8-S18-ESBL-04 and P. aeruginosa PAO1. Our findings highlight the necessity of expanding current biomonitoring indicators for wastewater treatment to optimize treatment strategies and mitigate the potential health risks posed by emerging pathogens. By addressing these research priorities, we can effectively mitigate risks and safeguard environmental safety and public health.

Complete mitochondrial DNA genome of banded cichlid Heros severus Heckel, 1840 (Perciformes: Cichlidae).

In this study, we sequenced the complete mitochondrial genome of Heros severus Heckel, 1840 (Perciformes, Cichlidae). This mitochondrial genome, consisting of 16,577 base pairs (bp), contains 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and 2 noncoding control regions (control region and origin of light-strand replication) as those found in other vertebrates. Control region, of 917 bp in length, is located between tRNAPro and tRNAPhe. Within the control region, typical conserved domains, such as the termination-associated sequence (TAS), central, and conserved sequence blocks domains were identified. The overall base composition of the heavy strand shows 27.6% of T, 26.3% of C, 29.3% of A, and 16.8% of G, with a slight A + T rich feature (56.9%). The complete mitogenome data provide useful genetic markers for the studies on the molecular identification, population genetics, phylogenetic analysis, and conservation genetics.

[Effects of matrine on airway inflammation and early airway remodeling in asthmatic mice].

OBJECTIVE: To observe the influence of matrine on airway inflammation and early airway remodeling in asthmatic mice. METHODS: Fifty BALB/c mice were randomly divided into 5 groups: a normal control group (A), an asthmatic group (B), a dexamethasone (DXM) group (C, 2 mg/kg), a high-dose matrine group (D, 50 mg/kg) and a low-dose matrine group (E, 25 mg/kg). The mice model of asthma in the B, C, D, and E groups was established by ovalbumin (OVA) intraperitoneal injections and aerosolization. Intra-gastric administration of different medications in C, D, E groups and 0.9% sodium chloride in B group were carried out 1 hour before provocation. 0.9% sodium chloride was used for intraperitoneal injection, aerosolization and intra-gastric administration in group A. The lung tissue slices were stained, and then the grade of inflammation around the wall of bronchi, mucous secretion, and the percentage of goblet-cells were counted. The areas of bronchial smooth muscle and of collagen deposition in airway wall were analyzed. The transcriptions and protein expressions of transforming growth factor-beta(1) (TGF-beta(1)) and connective tissue growth factor (CTGF) were measured respectively by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. RESULTS: In the A, B, C, D, E groups, the grades of inflammation were 1.5 (1, 2), 4 (4, 5), 2 (1, 3), 2 (2, 3), 3 (2, 3.3), respectively; the degrees of mucous secretions were 1.5 (1, 2), 5 (4, 6), 2 (1, 3), 2 (2.5, 4), 3 (3, 4), respectively. These airway inflammatory parameters in group B were significantly higher than in group A (chi(2) = 21.3, 22.6, P all < 0.01), while they were remarkably decreased in group C compared to group B (chi(2) = 13.3, 15.0, P all < 0.01). These parameters in group D and group E were also lower than those in group B (chi(2) = 9.1, 10.9; 9.8, 9.7; P all < 0.05). The percentage of goblet cells in airway epithelium was (1.7 +/- 0.5)%, (54.7 +/- 15.5)%, (20.4 +/- 5.9)%, (31.7 +/- 7.6)% and (36.2 +/- 10.8)%, respectively; it was significantly higher in group B than in group A (t = 12.0, P < 0.01), and remarkably lower in groups C and D than in group B (t = 7.7, 5.1, P all < 0.01), and lower in group E than in B group (t = 4.2, P < 0.05). In these 5 groups, the area of bronchial smooth muscle was (11.5 +/- 2.1) microm(2)/microm, (30.0 +/- 3.3) microm(2)/microm, (15.2 +/- 3.1) microm(2)/microm, (22.2 +/- 4.8) microm(2)/microm and (26.5 +/- 3.4) microm(2)/microm, respectively; it was significantly higher in group B than in group A (t = 11.4, P < 0.01), and remarkably lower in groups C, D and E than in group B (t = 9.1, 4.7, 2.2, P all < 0.01). The area of collagen deposition was (3.9 +/- 1.8) microm(2)/microm, (24.4 +/- 6.1) microm(2)/microm, (15.4 +/- 3.5) microm(2)/microm, (16.6 +/- 6.0) microm(2)/microm and (17.5 +/- 4.4) microm(2)/microm, respectively; it was also significantly higher in group B than in group A (t = 9.3, P < 0.01), and remarkably lower in groups C and D than in group B (t = 4.1, 3.5, P all < 0.01), and lower in group E than in B group (t = 3.2, P < 0.05). The mRNA levels of TGF-beta(1) were 160 +/- 25, 247 +/- 37, 174 +/- 23, 195 +/- 25 and 207 +/- 42, respectively, and those of CTGF were 86 +/- 8, 160 +/- 24, 94 +/- 10, 93 +/- 14 and 104 +/- 10, respectively in the 5 groups. The levels were remarkably increased in group B, as compared to group A (t = 6.1, 11.6, P all < 0.01), and the levels in groups C, D and E were remarkably decreased, as compared to group B, the difference being significant (t = 3.7, 2.7, 5.1; 10.6, 8.6, 10.3; P all < 0.01). The protein level of TGF-beta(1) in lung tissues was 21 +/- 5, 36 +/- 8, 26 +/- 5, 26 +/- 5 and 26 +/- 5, respectively, and that of CTGF was 15 +/- 4, 27 +/- 5, 21 +/- 4, 22 +/- 3 and 23 +/- 4, respectively in the 5 groups. The levels in B group were significantly increased, as compared to group A (t = 5.7, 6.4, P all < 0.01), and those in groups C and D were significantly decreased (t = 3.9, 3.9; 3.2, 2.8, P all < 0.01), and that in group E was also lower (t = 3.8, 2.5, P all < 0.05), as compared to group B. In all the groups, the protein levels of TGF-beta(1) and CTGF were positively correlated with the area of bronchial smooth muscle and with the area of collagen deposition (r = 0.435, 0.583, 0.522, 0.590, P all < 0.01). CONCLUSIONS: Matrine inhibited airway inflammation and early airway remodeling in asthmatic mice. The signal transduction of TGF-beta(1) and CTGF maybe involved.

Kisspeptin­13 inhibits bleomycin­induced pulmonary fibrosis through GPR54 in mice.

Kisspeptin (KP) is an amidated neurohormone that is encoded by the KiSS­1 metastasis suppressor (KISS1) gene and serves as the endogenous ligand for G protein­coupled receptor 54 (GPR54). KP is involved in the regulation of several biological functions, such as reproduction, cancer and atherogenesis. Recent data suggested that KP may induce atherosclerotic plaque progression and instability, which may be reversed by the GPR54 antagonist KP­234. Despite the KISS1 gene being previously reported as a downstream target of the classic transforming growth factor (TGF)/Smad2 signaling pathway, its role in fibrosis remains elusive. The purpose of the present study was to evaluate the role of KP­13 (a product of the KISS1 gene) in a bleomycin (BLM)­induced idiopathic pulmonary fibrosis model. Lung tissue samples were evaluated by quantitative PCR analysis, western blotting and ELISA. Daily intraperitoneal administration of KP­13 significantly ameliorated body weight loss, histopathological lung abnormalities and pulmonary collagen deposition induced by BLM. Furthermore, KP­13 downregulated the expression levels of tumor necrosis factor­α, TGF­ß, collagen type I α1, actin α2 and matrix metalloproteinase 2 in BLM­treated lungs compared with BLM group. Notably, the production of α­smooth muscle actin in lung tissues, as well as the pulmonary levels of TGF­ß1 and phosphorylated­Smad2/3, was reduced following treatment with KP­13. The anti­fibrotic effects of KP­13 were reversed by KP­234 (an antagonist of GPR54), but not by Cetrorelix (an antagonist of the gonadotropin­releasing hormone receptor). Furthermore, apoptosis­related proteins, such as Bax and caspase­3, were decreased, whereas Bcl­2 was markedly increased as determined by western blotting. Collectively, these data suggested that the KP/GPR54 signaling pathway may be a promising target for the treatment of idiopathic pulmonary fibrosis.

[A study on oxidative DNA damage and lipid peroxidation in patients with tuberculous pleurisy].

OBJECTIVE: To explore the levels of mononuclear cell oxidative DNA damage and lipid peroxidation in patients with tuberculous pleurisy. METHODS: The mononuclear cell DNA damages in pleural effusion and peripheral blood of 28 patients with tuberculous pleurisy and in peripheral blood of 25 healthy persons were detected by single cell gel electrophoresis (comet%). The levels of total antioxidative capacity (TAC) in supernatant of pleural effusion and blood plasma of 28 patients and in blood plasma of 25 healthy persons were measured by o-phenanthroline colorimetric analysis. The contents of malondialdehyde (MDA) of blood plasma from 28 patients and from 25 healthy persons were measured by thiobarbituric acid colorimetric analysis. The differences between groups were analyzed by t test. RESULTS: The comet percentage of mononuclear cells in pleural effusion from patients with tuberculous pleurisy was (41.3 +/- 14.5)%, which was significantly higher than that in the peripheral blood (21.2 +/- 4.2)% (P < 0.01); The level of TAC in supernatant of pleural effusion was (5 172 +/- 1 195) U/L, which was lower than that in the blood plasma (8 656 +/- 1 592) U/L (P < 0.01). There was a negative correlation between the comet percentage of mononuclear cells and the level of TAC (r = -0.425, P < 0.05) in pleural effusion. The comet percentage of mononuclear cells in peripheral blood and the content of MDA in blood plasma from patients were (21.2 +/- 4.2)% and (8.25 +/- 1.37) micro mol/L, which were significantly higher than those from the controls (8.9 +/- 3.7)% and (4.46 +/- 0.93) micro mol/L respectively (P < 0.01); The level of TAC in blood plasma was (8 656 +/- 1 592) U/L, which was lower than that from the controls (10 610 +/- 1 399) U/L (P < 0.01). There were negative correlations between the level of TAC in blood plasma and the comet percentage of mononuclear cells in peripheral blood or the content of MDA in blood plasma (r = -0.438, -0.413, P < 0.05), and there was a positive correlation between the comet percentage of mononuclear cells in peripheral blood and the content of MDA in blood plasma (r = 0.899, P < 0.01). CONCLUSION: There is oxidation/antioxidation imbalance in patients with tuberculous pleurisy, namely oxidative stress, with inadequate total antioxidative capacity, particularly in the diseased pleural cavity.