Hirudin ameliorates myocardial ischemia-reperfusion injury in a rat model of hemorrhagic shock and resuscitation: roles of NLRP3-signaling pathway.

Severe hemorrhage shock and resuscitation (HSR) has been reported to induce myocardial ischemia-reperfusion injury (MIRI), resulting in a poor prognosis. Hirudin, an effective thrombin inhibitor, can offer protection against MIRI. This study aimed to determine if hirudin administration ameliorates HSR-induced MIRI and the underlying mechanism. A rat model of HSR was established by bleeding rats to a mean arterial blood pressure of 30-35 mmHg for 45 min and then resuscitating them with all the shed blood through the left femoral vein. After HSR, 1 mg/kg of hirudin was administrated immediately. At 24 h after HSR, the cardiac injury was assessed using serum CK-MB, cTnT, hematoxylin-eosin (HE) staining, echocardiography, M1-polarized macrophages, and pyroptosis-associated factors, including cleaved caspase-1, Gasdermin D (GSDMD) N-terminal, IL-1ß, and IL-18 were measured by immunofluorescence and western blot assays. Nigericin, a unique agonist, was utilized to evaluate the responsibilities of NLRP3 signaling. Under the HSR condition, rats exhibited a significant increase in myocardial injury score, an elevation of serum cTnT, CK-MB levels, an aggrandization of M1-polarized macrophages, an upregulation of pyroptosis-associated factors, including cleaved caspase-1, GSDMD N-terminal, IL-1ß, and IL-18, but a significant decrease in left ventricular ejection fraction (EF%) and a reduction of left ventricular fractional shortening (FS%), while hirudin administration partially restored the changes. However, the NLRP3 agonist nigericin reversed the cardioprotective effects of hirudin. We determined the cardioprotective effects of hirudin against HSR-induced MIRI. The mechanism may involve the inhibition of NLRP3-induced pyroptosis.

NLRP3-GABA signaling pathway contributes to the pathogenesis of impulsive-like behaviors and cognitive deficits in aged mice.

BACKGROUND: Perioperative neurocognitive disorders (PND), such as delirium and cognitive impairment, are commonly encountered complications in aged patients. The inhibitory neurotransmitter γ-aminobutyric acid (GABA) is aberrantly synthesized from reactive astrocytes following inflammatory stimulation and is implicated in the pathophysiology of neurodegenerative diseases. Additionally, the activation of NOD-like receptor protein 3 (NLRP3) inflammasome is involved in PND. Herein, we aimed to investigate whether the NLRP3-GABA signaling pathway contributes to the pathogenesis of aging mice's PND. METHODS: 24-month-old C57BL/6 and astrocyte-specific NLRP3 knockout male mice were used to establish a PND model via tibial fracture surgery. The monoamine oxidase-B (MAOB) inhibitor selegiline (1 mg/kg) was intraperitoneally administered once a day for 7 days after the surgery. PND, including impulsive-like behaviors and cognitive impairment, was evaluated by open field test, elevated plus maze, and fear conditioning. Thereafter, pathological changes of neurodegeneration were explored by western blot and immunofluorescence assays. RESULTS: Selegiline administration significantly ameliorated TF-induced impulsive-like behaviors and reduced excessive GABA production in reactive hippocampal astrocytes. Moreover, astrocyte-specific NLRP3 knockout mice reversed TF-induced impulsive-like and cognitive impairment behaviors, decreased GABA levels in reactive astrocytes, ameliorated NLRP3-associated inflammatory responses during the early stage, and restored neuronal degeneration in the hippocampus. CONCLUSIONS: Our findings suggest that anesthesia and surgical procedures trigger neuroinflammation and cognitive deficits, which may be due to NLRP3-GABA activation in the hippocampus of aged mice.

Spautin-1 Protects Against Mild TBI-Induced Anxiety-Like Behavior in Mice via Immunologically Silent Apoptosis.

Anxiety is reportedly one of the most common mental changes after traumatic brain injury (TBI). Perineuronal nets (PNNs) produced by astrocytes in the lateral hypothalamus (LHA) that surround gamma-aminobutyric acid-ergic (GABAergic) neurons have been associated with anxiety. The potent anti-tumor effects of Spautin-1, a novel autophagy inhibitor, have been documented in malignant melanoma; moreover, the inhibition of autophagy is reported to mitigate anxiety disorders. However, little is known about the ability of spautin-1 to alleviate anxiety. In this study, we sought to investigate whether spautin-1 could alleviate anxiety-like behaviors post-TBI by reducing the loss of PNNs in the LHA. A mild TBI was established in mice through Feeney's weight-drop model. Then, Spautin-1 (20 mmol/2 µl) was immediately administered into the left lateral ventricle. Behavioral and pathological changes were assessed at 24 h, 7 days, 30 days, 31 days and 32 days after TBI by the neurological severity scores (NSS), open field test (OFT), elevated plus-maze (EPM) test, western blot, immunofluorescence assays and electron microscopy. Spautin-1 significantly reversed TBI-induced decreased time in the central zone during OFT and in the open-arm during the EPM test. Spautin-1 also increased PNNs around GABAergic neurons indicated by WFA- plus GAD2- positive A2-type astrocytes and attenuated M1-type microglia in the LHA 32 days after TBI compared to TBI alone. Moreover, compared to mice that only underwent TBI, spautin-1 downregulated autophagic vacuoles, abnormal organelles, the expression of Beclin 1, USP13, phospho-TBK1, and phospho-IRF3 and upregulated the levels of cleaved caspase-3, -7 and -9, but failed to increase TUNEL-positive cells in the LHA at 24 h. Spautin-1 alleviated anxiety-like behavior in mice exposed to mild TBI; this protective mechanism may be associated with decreased PNNs loss around GABAergic neurons via immunologically silent apoptosis induced by the caspase cascade.

Abdominal surgery plus sevoflurane exposure induces abnormal emotional changes and cognitive dysfunction in aged rats.

BACKGROUND: Cognitive impairment, which includes perioperative psychological distress and cognitive dysfunction, can be determined by preoperative and post-operative neuropsychological tests. Several mechanisms have been proposed regarding the two-way communication between the immune system and the brain after surgery. We aimed to understand the mechanisms underlying perioperative neurocognitive disorders (PND) in elderly rats using an experimental abdominal surgery model. METHODS: 24-month-old SD rats were exposed to the abdominal surgery model (AEL) under 3% anesthesia. On day 15 and day 30 post-surgery, fractional anisotropy (FA) using diffusion kurtosis imaging (DKI) was measured. From day 25 to day 30 post-surgery, behavioral tests, including open field test (OFT), Morris water maze (MWM), novel object recognition (NOR), force swimming test (FST), and elevated plus maze (EPM), were performed. Then, the rats were euthanized to perform pathological analysis and western blot measurement. RESULTS: The rats exposed to AEL surgical treatment demonstrated significantly decreased time crossing the platform in the MWM, decreased recognition index in the NOR, reduced time in the open arm in the EPM, increased immobility time in the FST, and increased number of crossings in the OFT. Aged rats, after AEL exposure, further demonstrated decreased FA in the mPFC, nucleus accumbens (NAc), and hippocampus, together with reduced MAP2 intensity, attenuation of GAD65, VGlut2, CHAT, and phosphorylated P38MAPK expression, and increased reactive astrocytes and microglia. CONCLUSIONS: In this study, the aged rats exposed to abdominal surgery demonstrated both emotional changes and cognitive dysfunction, which may be associated with neuronal degeneration and reduced phosphorylated P38MAPK.

IL-18BP Alleviates Anxiety-Like Behavior Induced by Traumatic Stress via Inhibition of the IL-18R-NLRP3 Signaling Pathway in a Mouse Model of Hemorrhagic Shock and Resuscitation.

Psychological distress and posttraumatic stress, including anxiety, severely influence life quality. Previously, we reported that interleukin-18 (IL-18) was involved in pyroptosis-induced emotional changes in a rodent model of hemorrhagic shock and resuscitation (HSR). Here, we aimed to continue our investigation on the role of IL-18 binding protein (IL-18BP), which exhibits excellent anti-inflammatory effects as an IL-18 negative regulator. Mice were administered with an intraperitoneal injection of IL-18BP after HSR exposure and anxiety-like behavior was examined using the open-field test and elevated plus maze test. Moreover, the following variables post-HSR were measured: (1) the activation of astrocytes; (2) pyroptosis-associated factors including cleaved caspase-1, GSDMD, IL-18; (3) the roles of IL-18 receptor (IL-18R)-NOD-like receptor pyrin domain-containing-3 (NLRP3) signal with the application of the NLRP3 specific agonist or astrocyte-specific NLRP3 knockout mice. IL-18BP administration remarkably alleviated HSR-induced anxiety-like behavior, astrocytic activation, and increases in pyroptosis-associated factors, while NLRP3 agonist nigericin partially reversed IL-18BP-induced neuroprotective effects. Astrocyte-specific NLRP3 knockout mice exhibited relatively less anxiety-like behavior. Similarly, IL-18BP exhibited an anti-pyroptosis effect in astrocytes in an in vitro model of low oxygen-glucose deprivation. These findings offer unique perspectives on HSR-induced posttraumatic stress and indicate that inhibition of IL-18R-NLRP3 signal via IL-18BP can attenuate astrocytic activation and pyroptosis, broadening the therapeutic landscape for patients with psychological distress and posttraumatic stress.

Subanesthetic dose of S-ketamine improved cognitive dysfunction via the inhibition of hippocampal astrocytosis in a mouse model of post-stroke chronic stress.

Post-stroke chronic stress (PSCS) is generally associated with the poorer recovery and more pronounced cognitive dysfunction. Recent evidence has implied that S-ketamine can reduce suicidal ideation in treatment-resistant depression. In this current study, we aimed to investigate whether the administration of S-ketamine ameliorated cognitive deficits under PSCS conditions, which was established by a model combining middle cerebral artery occlusion (MCAO) and chronic restraint stress. Our data suggested that mice exposed to PSCS exhibited depression-like behavior and cognitive impairment, which coincided with astrocytosis as indicated by increased GFAP-positive cells and impairment of long-time potentiation (LTP) in the hippocampal CA1. Subanesthetic doses (10 mg/kg) of S-ketamine have significantly mitigated depression-like behaviors, cognitive deficits and LTP impairment, reduced astrocytosis, excessive GABA, and inflammatory factors, including NLRP3 and IL-18 in astrocytes in the CA1. Besides, neuroprotective effects induced by S-ketamine administration were found in vitro but could be partially reversed by an agonist of the NLRP3 nigericin. Our current data also suggests that the subanesthetic doses of S-ketamine improved cognitive dysfunction via the inhibition of hippocampal astrocytosis in a mouse model of PSCS.

Characterization and Antibacterial Properties of Polyetherketoneketone Coated with a Silver Nanoparticle-in-Epoxy Lining.

Polyetherketoneketone (PEKK) is an alternative material for use in removable partial denture frameworks; these frameworks must exhibit antibacterial properties to reduce the risk of periodontal disease. In the present study, silver nanoparticles (AgNPs) were synthesized via the reduction of silver nitrate with sodium borohydride in a solution containing polyvinyl pyrrolidone (PVP). Transmission electron microscope images and dynamic light scattering confirmed that metallic nanoparticles had been created with an average size of 32 nm. Furthermore, the coating of the PEKK polymeric substrate with 0.5% AgNPs was carried out using an epoxy resin lining at room temperature. Fourier transform infrared (FTIR) spectra confirmed the successful transfer of the AgNP-in-resin lining onto the polymeric substrate. Scanning electron microscopy and atomic force microscopy confirmed that the AgNPs had been uniformly deposited onto the PEKK specimens. Finally, the antibacterial activity of the specimens was tested against Porphyromonas gingivalis. An inhibition zone of 22.5 mm and an antibacterial rate of 83.47% were found for the PEKK coated with 0.5% AgNPs (0.5% Ag-PEKK) compared to an untreated polyetheretherketone (PEEK) substrate, evidencing that 0.5% Ag-PEKK has potential antibacterial properties for implant applications.

[Structure and Functional Diversity of Bacterial Community in Rhizosphere Soil of Typical Vegetation in the Riparian Zone Along the Downstream of Songhua River].

The aim of this study was to provide a reference for the riparian zone with protection and ecological restoration by analyzing the differences in typical vegetation (Phragmites communis, Populus tomentosa, Salix sungkianica, and Carex schmidtii) rhizosphere bacterial communities and their functions and identifying the potential of different types of vegetation to restore the damaged riparian zone in Songhua River. The 16S rRNA of rhizosphere soil bacteria in the four typical vegetation types of the riparian zone along the downstream of the Songhua River was sequenced using the Illumina MiSeq PE300 high-throughput sequencing platform. The community diversity, functional differences, and influencing factors of rhizosphere soil bacteria for different vegetation types were analyzed. The results showed that the Ace index, Chao1 index, and Shannon index of soil bacterial diversity in P.communis were significantly higher than those of P.tomentosa (P<0.05), and there was no significant difference between the above two types of vegetation and S. sungkianica and C.schmidtii. There were significant differences between the soil bacterial community structure of P. tomentosa and that of the three other vegetation types (P<0.05). The soil bacterial community structures of S.sungkianica, C.schmidtii, and P.communis were similar. Bacteria in the rhizosphere soil of the four typical vegetation types could be divided into 38 phyla. Acidobacteria, Actinobacteria, Chloroflexi, Verrucomicrobia, and Proteobacteria were the dominant phyla (relative abundance>5%), and the Nitrospirae, Gemmatimonadetes, Bacteroidetes, Firmicutes, and Rokubacteria of bacteria had a relative abundance greater than 1%. The bacterial community in the rhizosphere soil of the four typical vegetation types had 6 primary metabolic pathways and 43 secondary metabolic pathways, including 14 types of main secondary metabolic pathways (relative abundance>1%). Diversity in rhizosphere soil bacterial communities of different vegetation types was significantly influenced by the C/N ratio, soil pH, and moisture content. Hence, the effects of different vegetation types in repairing the degraded riparian zone were different, and wetland vegetation (S.sungkianica and C.schmidtii) was conducive to the improvement in soil bacterial diversity and soil ecosystem functions.

Dual RNA-seq provides novel insight into the roles of dksA from Pseudomonas plecoglossicida in pathogen-host interactions with large yellow croakers ( Larimichthys crocea).

Pseudomonas plecoglossicida is a rod-shaped, gram-negative bacterium with flagella. It causes visceral white spot disease and high mortality in Larimichthys crocea during culture, resulting in serious economic loss. Analysis of transcriptome and quantitative real-time polymerase chain reaction (PCR) data showed that dksA gene expression was significantly up-regulated after 48 h of infection with Epinephelus coioides (log 2FC=3.12, P<0.001). RNAi of five shRNAs significantly reduced the expression of dksA in P. plecoglossicida, and the optimal silencing efficiency was 96.23%. Compared with wild-type strains, the symptoms of visceral white spot disease in L. crocea infected with RNAi strains were reduced, with time of death delayed by 48 h and mortality reduced by 25%. The dksA silencing led to a substantial down-regulation in cellular component-, flagellum-, and ribosome assembly-related genes in P. plecoglossicida, and the significant up-regulation of fliC may be a way in which virulence is maintained in P. plecoglossicida. The GO and KEGG results showed that RNAi strain infection in L. crocea led to the down-regulation of inflammatory factor genes in immune-related pathways, which were associated with multiple immune response processes. Results also showed that dksA was a virulence gene in P. plecoglossicida. Compared with the wild-type strains, RNAi strain infection induced a weaker immune response in L. crocea.

Development of Self-Healable Organic/Inorganic Hybrid Materials Containing a Biobased Copolymer via Diels-Alder Chemistry and Their Application in Electromagnetic Interference Shielding.

In this study, a novel biobased poly(ethylene brassylate)-poly(furfuryl glycidyl ether) copolymer (PEBF) copolymer was synthesized and applied as a structure-directing template to incorporate graphene and 1,1'-(methylenedi-4,1-phenylene)bismaleimide (BMI) to fabricate a series of self-healing organic/inorganic hybrid materials. This ternary material system provided different types of diene/dienophile pairs from the furan/maleimide, graphene/furan, and graphene/maleimide combinations to build a crosslinked network via multiple Diels-Alder (DA) reactions and synergistically co-assembled graphene sheets into the polymeric matrix with a uniform dispersibility. The PEBF/graphene/BMI hybrid system possessed an efficient self-repairability for healing structural defects and an electromagnetic interference shielding ability in the Ku-band frequency range. We believe that the development of the biobased self-healing hybrid system provides a promising direction for the creation of a new class of materials with the advantages of environmental friendliness as well as durability, and shows potential for use in advanced electromagnetic applications.

Combining the Photocatalysis and Absorption Properties of Core-Shell Cu-BTC@TiO2 Microspheres: Highly Efficient Desulfurization of Thiophenic Compounds from Fuel.

A core-shell Cu-benzene-1,3,5-tricarboxylic acid (Cu-BTC)@TiO2 was successfully synthesized for photocatalysis-assisted adsorptive desulfurization to improve adsorptive desulfurization (ADS) performance. Under ultraviolet (UV) light irradiation, the TiO2 shell on the surface of Cu-BTC achieved photocatalytic oxidation of thiophenic S-compounds, and the Cu-BTC core adsorbed the oxidation products (sulfoxides and sulfones). The photocatalyst and adsorbent were combined using a distinct core-shell structure. The morphology and structure of the fabricated Cu-BTC@TiO2 microspheres were verified by scanning electron microscopy, high-resolution transmission electron microscopy, energy-dispersive x-ray spectroscopy, X-ray powder diffraction, nitrogen adsorption-desorption and X-ray photoelectron spectroscopy analyses. A potential formation mechanism of Cu-BTC@TiO2 is proposed based on complementary experiments. The sulfur removal efficiency of the microspheres was evaluated by selective adsorption of benzothiophene (BT) and dibenzothiophene (DBT) from a model fuel with a sulfur concentration of 1000 ppmw. Within a reaction time of 20 min, the BT and DBT conversion reached 86% and 95%, respectively, and achieved ADS capacities of 63.76 and 59.39 mg/g, respectively. The BT conversion and DBT conversion obtained using Cu-BTC@TiO2 was 6.5 and 4.6 times higher, respectively, than that obtained using Cu-BTC. A desulfurization mechanism was proposed, the interaction between thiophenic sulfur compounds and Cu-BTC@TiO2 microspheres was discussed, and the kinetic behavior was analyzed.

Self-Assembled pH and Redox Dual Responsive Carboxymethylcellulose-Based Polymeric Nanoparticles for Efficient Anticancer Drug Codelivery.

To face the growing demand of polymeric nanoparticles with biocompatibility and a drug release profile, in this work, a novel carboxymethylcellulose-based pH and redox dual-responsive polymeric nanoparticle, carboxymethyl cellulose-dithiopropionate hydrazide-8arm-polyethylene glycol-pterostilbene/10-hydroxy camptothecin (CTPP/HCPT), was prepared for efficient drug codelivery. These well-dispersed CTPP/HCPT NPs were prepared with a dimension of around 144 nm and exhibited high binary drug loading capacity and good biocompatibility. The biggest advantage of this design is that these nanoparticles can rapidly release the drug payload responding to intracellular acidic or reductive stimuli, while maintaining sufficient stability under normal physiological conditions. The in vitro drug release study revealed that the HCPT payload released from nanoparticles in a weakly acidic environment with 10 mM reductive glutathione was about 74.8%, which was 3.8-fold higher than under normal physiological conditions (∼19.6%). Further in vitro and in vivo investigation demonstrated that such dual-responsive CTPP/HCPT NPs could potently kill cancer cells and suppress tumor growth with lower adverse effects. All these results suggested that CTPP/HCPT NPs were suitable as potential and effective candidates for cancer therapy.

Correlation of FOXC1 protein with clinicopathological features in serous ovarian tumors.

Transcriptional factor FOXC1 has been demonstrated to play a key role in embryogenesis in animal studies and may participate in tumorigenesis. However, the specific function of this gene in ovarian tumors has not been fully determined. In this study, potential correlations between FOXC1 expression and clinicopathological features of serous ovarian tumors were investigated. FOXC1 expression was analyzed in SKOV-3 and HO-8910 cell lines and serous ovarian tumor tissues. A significant correlation was observed between FOXC1 protein expression and pathological subtype as well as FIGO stage (P<0.05) in serous ovarian tumors in our retrospective study. No significant association was revealed between FOXC1 protein expression and the clinicopathological factors of age, histological grade and volume of ascites (P>0.05). The results suggest that high expression of FOXC1 protein may serve as a marker for benign serous ovarian tumors and a suggest a trend towards good prognosis.

[Comparative study on effect of acupuncture and lidocaine block for lumbar myofascial pain syndrome].

OBJECTIVE: To observe the clinical efficacy of acupuncture at Jiaji (EX-B 2) points mainly for lumbar myofascial pain syndrome (MPS). METHODS: Sixty-six cases of MPS were randomized into an acupuncture group and a lidocaine group, 33 cases in each group. The acupuncture group was treated with acupuncture at Jiaji (EX-B 2) points combined with needling local myofascial trigger points (MTrP), and the lidocaine group was treated with local block at trigger points with lidocaine injection. The treatment was given once every 2 days. After three and five times of the treatment, the simplified McGill scale, Oswestry disability index (ODI) and pressure-pain threshold were assessed to compare the therapeutic effects between the two groups. RESULTS: After treatment, the scores of simplified McGill and ODI of two groups were obviously reduced while the score of pressure-pain threshold was obviously increased (all P < 0.01). After three and five times of the treatment, there were no significant differences in above scores between the two groups (all P > 0.05). CONCLUSION: Acupuncture at Jiaji (EX-B 2) points combined with needling MTrP is an effective and safe therapy for lumbar MPS, the therapeutic effect is equal to lidocaine block.

The antitumor effect of the toll-like receptor 3 ligand polyinosinic-cytidylic acid as an adjuvant.

Although polyinosinic-polycytidylic acid (poly(I:C)) has been applied in tumor immunity as a Toll-like receptor 3 (TLR3) ligand, the interaction between poly(I:C) and TLR3 is still unclear, as are the mechanisms underlying the antitumor effect of poly(I:C). Our aim was to investigate the interaction between poly(I:C) and TLR3, as well as the mechanisms underlying the antitumor effect of poly(I:C). NK92 cells were maintained in medium (untreated group), or medium containing E7(44-62) (E7 group) or E7(44-62)+poly(I:C) (poly(I:C)/E7 group), and we measured the expression of TLR3 mRNA, p-p65, and IκB-α protein. The cells were first incubated in medium alone or medium containing TLR3 monoclonal antibody, and then in medium containing poly(I:C)/E7. Finally, we measured the level of interferon-beta (INF-ß) in the supernatant and determined the tumor cell-killing effect of the NK92 cells. At 1 h, the expression of TLR3 mRNA in the poly(I:C)/E7 group was markedly higher than that in the untreated and E7 groups (P < 0.05). When compared with the poly(I:C)/E7 group, the expression of IκB-α was dramatically increased in the E7 and untreated groups, and the expression of p-p65 was dramatically decreased in the E7 and untreated groups (all P < 0.05). At 24 h, INF-ß content and tumor cell-killing activity in the poly(I:C)/E7 group were markedly higher than those in the untreated group (P < 0.001, <0.05, respectively). Treatment with TLR3 monoclonal antibody significantly inhibited poly(I:C)/E7-induced INF-ß secretion and tumor cell-killing activity in NK92 cells (P < 0.001, <0.05, respectively). The interaction between poly(I:C) and TLR3 plays an important role in the antitumor immunity of NK92 cells. In addition, the interaction between poly(I:C) and TLR3 increases INF-ß expression, which may be attributed to the activation of NFκB.