ObjectiveTo investigate the mechanisms by which Huanglian Jiedutang （HLJDT） modulates microglial （MG） phenotypes through the sialic acid-binding Ig-like lectin 2 （SIGLEC2/CD22）/Src-homology-2-domain-containing protein tyrosine phosphatase-1 （SHP-1）/phosphorylated protein kinase B （p-Akt） signaling pathway， thereby promoting myelin repair and alleviating agitation-like behaviors in vascular dementia （VAD）. MethodsSixty C57BL/6J mice were randomly assigned to a sham （normal） group， model group， HLJDT low-， medium-， and high-dose groups （2.5， 5， and 10 g·kg^-1·d^-1）， and a risperidone group （2 mg·kg^-1·d^-1）， with 10 mice per group. VAD was induced by bilateral common carotid artery stenosis （BCAS）. From day 42， mice received drug interventions for 2 weeks. Agitation-like behaviors were assessed using the resident-intruder test. After behavioral testing， ventrolateral part of the ventromedial hypothalamus （VMHvl） tissues were collected. Western blot was used to measure protein levels of myelin oligodendrocyte glycoprotein （MOG）， myelin basic protein （MBP）， proteolipid protein （PLP）， inducible nitric oxide synthase （iNOS）， arginase-1 （Arg1）， CD86， CD206， and CD22， SHP-1， and p-Akt. Immunofluorescence was used to evaluate myelin-associated glycoprotein （MAG） intensity and the proportion of iNOS⁺/ionized calcium-binding adapter molecule 1 （Iba1）⁺ cells. ELISA was used to detect tumor necrosis factor-α （TNF-α）， interleukin （IL）-6， and IL-1β. ResultsCompared with the normal group， the model group exhibited markedly increased biting and aggressive behaviors and shortened attack latency （P<0.01）. MOG， MBP， and PLP protein levels and MAG fluorescence intensity were significantly reduced （P<0.05， P<0.01）. INOS and CD86 expression and TNF-α， IL-6， and IL-1β levels were significantly elevated （P<0.01）. CD22 and SHP-1 expression increased significantly （P<0.01）， whereas p-Akt expression decreased （P<0.01）. Compared with the model group， the medium- and high-dose HLJDT groups and the risperidone group showed markedly reduced biting and aggression （P<0.05， P<0.01） and prolonged attack latency （P<0.01）. MOG， MBP， and PLP levels and MAG fluorescence intensity were significantly increased （P<0.05， P<0.01）. INOS， CD86， TNF-α， IL-6， and IL-1β levels decreased significantly （P<0.05， P<0.01）. CD22 and SHP-1 expression decreased， while p-Akt expression increased significantly （P<0.05， P<0.01）. ConclusionHLJDT may modulate CD22/SHP-1/p-Akt signaling in the VMHvl， promote the shift of MG toward an anti-inflammatory and phagocytic phenotype， enhance myelin repair， and improve agitation-like behaviors in VAD mice.

ObjectiveTo investigate the ameliorative effects of quercetin on neuropsychiatric symptoms associated with vascular dementia （VaD） and to elucidate the molecular mechanism， specifically whether quercetin inhibits pro-inflammatory activation of microglia by modulation of the receptor-interacting serine/threonine-protein kinase 1 （RIPK1）/NOD-like receptor protein 3 （NLRP3）/Caspase-1 signaling pathway， thereby promoting myelin repair in the medial prefrontal cortex （mPFC）. MethodsA C57BL/6J mouse model of VaD with neuropsychiatric symptoms was established by bilateral common carotid artery stenosis （BCAS） combined with chronic restraint stress （CRS）. Mice were randomly divided into a sham group， a model group， low-dose， medium-dose， and high-dose quercetin groups （30， 60， 120 mg·kg^-1·d^-1）， and a fluoxetine group （10 mg·kg^-1·d^-1）. After intervention， depressive- and anxiety-like behaviors were assessed by the sucrose preference test （SPT）， forced swim test （FST）， open field test （OFT）， and elevated plus maze （EPM）. mPFC tissue was collected. Immunofluorescence （IF） was used to detect myelin basic protein （MBP） expression and microglial morphology. Western blot was used to measure the protein level of MBP， myelin oligodendrocyte glycoprotein （MOG）， myelin-associated glycoprotein （MAG）， inducible nitric oxide synthase （iNOS）， CD86， RIPK1， phosphorylated RIPK1 （Ser166）， NLRP3， and Caspase-1. Enzyme-linked immunosorbent assay （ELISA） was used to determine the level of tumor necrosis factor-alpha （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β）. ResultsCompared with the sham group， the model group exhibited significant depressive- and anxiety-like behaviors （P<0.01）， significantly decreased protein expression of MBP， MOG， and MAG in the mPFC （P<0.01）， activated microglia （characterized by enlarged cell bodies， reduced protrusions， and upregulated iNOS and CD86 expressions， P<0.01）， and significantly elevated p-RIPK1/RIPK1 ratio， NLRP3， Caspase-1 protein expression， and level of TNF-α， IL-6， and IL-1β （P<0.01， P<0.05）. Compared with the model group， the quercetin treatment （especially at medium and high doses） significantly ameliorated these behavioral abnormalities （P<0.05， P<0.01）， increased the expression of MBP （protein and fluorescence intensity）， MOG， and MAG in the mPFC （P<0.05， P<0.01）， suppressed excessive microglial activation （characterized decreased cell bodies， increased protrusions， and downregulated iNOS and CD86 expressions， P<0.01）， and significantly reduced the p-RIPK1/RIPK1 ratio， NLRP3， Caspase-1 protein expression， and inflammatory cytokine levels （P<0.01）. ConclusionQuercetin effectively alleviates neuropsychiatric symptoms in VaD mice. Its mechanism may be associated with the inhibition of microglial inflammatory activation mediated by the RIPK1/NLRP3/Caspase-1 signaling pathway， thereby promoting myelin repair in the mPFC region.

AIM:To compare the therapeutic effects of liproxstatin-1(LIP-1)and N-acetylcysteine(NAC)in bleomycin(BLM)-induced pulmonary fibrosis.METHODS:The mice were randomly divided into the control,model(BLM),BLM+NAC,BLM+LIP-1,NAC and LIP-1 groups.The BLM+NAC and BLM+LIP-1 groups were treated with NAC by intratracheal drip and LIP-1 by intraperitoneal injection,respectively,1 day before BLM tracheal instillation.The other groups received intraperitoneal injection of the same volume of co-solvent and intragastric instillation of saline.Fourteen days after a BLM challenge,the degree of lung fibrosis and the expression levels of alveolar epithelial cell mark-ers and ferroptosis-related molecules were assessed in each group.RESULTS:LIP-1 treatment more significantly im-proved the BLM-induced decrease in body weight(P＜0.01)and survival rate in mice compared with NAC.LIP-1 more significantly reduced the severity of pulmonary fibrosis and improved collagen deposition compared with NAC.LIP-1 also more significantly alleviated alveolar structural disruption,and more significantly inhibited the decrease in the alveolar epi-thelial cell markers podoplanin and surfactant protein C,as well as epithelial-mesenchymal transition,compared with NAC.LIP-1 was a more potent inhibitor of the BLM-induced increase in ferroptosis and its related molecule Heme oxygen-ase-1 than NAC.CONCLUSION:LIP-1 treatment is more effective than NAC in alleviating pulmonary fibrosis.Mecha-nistically,this finding may be related to the ability of LIP-1 to inhibit ferroptosis in alveolar epithelial cells.This study pro-vides new insights into the treatment of pulmonary fibrosis and lays the foundation for the clinical application of LIP-1.

AIM:To compare the therapeutic effects of liproxstatin-1(LIP-1)and N-acetylcysteine(NAC)in bleomycin(BLM)-induced pulmonary fibrosis.METHODS:The mice were randomly divided into the control,model(BLM),BLM+NAC,BLM+LIP-1,NAC and LIP-1 groups.The BLM+NAC and BLM+LIP-1 groups were treated with NAC by intratracheal drip and LIP-1 by intraperitoneal injection,respectively,1 day before BLM tracheal instillation.The other groups received intraperitoneal injection of the same volume of co-solvent and intragastric instillation of saline.Fourteen days after a BLM challenge,the degree of lung fibrosis and the expression levels of alveolar epithelial cell mark-ers and ferroptosis-related molecules were assessed in each group.RESULTS:LIP-1 treatment more significantly im-proved the BLM-induced decrease in body weight(P＜0.01)and survival rate in mice compared with NAC.LIP-1 more significantly reduced the severity of pulmonary fibrosis and improved collagen deposition compared with NAC.LIP-1 also more significantly alleviated alveolar structural disruption,and more significantly inhibited the decrease in the alveolar epi-thelial cell markers podoplanin and surfactant protein C,as well as epithelial-mesenchymal transition,compared with NAC.LIP-1 was a more potent inhibitor of the BLM-induced increase in ferroptosis and its related molecule Heme oxygen-ase-1 than NAC.CONCLUSION:LIP-1 treatment is more effective than NAC in alleviating pulmonary fibrosis.Mecha-nistically,this finding may be related to the ability of LIP-1 to inhibit ferroptosis in alveolar epithelial cells.This study pro-vides new insights into the treatment of pulmonary fibrosis and lays the foundation for the clinical application of LIP-1.

This work aims to explore the effect of glycolysis on the replication of porcine reproductive and respiratory syndrome virus (PRRSV) in porcine alveolar macrophages (PAMs). The changes of glucose metabolism, PRRSV protein levels, PRRSV titers, and the relative expression levels of genes and proteins in PAMs were analyzed by ELISA, qPCR, virus titration, and Western blotting after PRRSV infection. The effect of hypoxia-inducible factor-1α (HIF-1α) on PRRSV replication was subsequently assessed by specific siRNAs targeting to HIF-1α. The results showed that PRRSV infection enhanced glycolysis, elevated the levels of glucose uptake and lactate in the supernatant (P<0.05 and 0.01, respectively), reduced ATP production (P<0.05), and up-regulated the expression of hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2, 6-biphosphatase 3 (PFKFB3), and pyruvate kinase isozyme type M2 (PKM2) in PAMs (P<0.05 and 0.01, respectively). Glycolysis inhibitors down-regulated the expression of PRRSV proteins and reduced virus titers (P<0.01). The knockdown of HIF-1α by siRNAs inhibited glycolysis and lowered PRRSV titers (P<0.05). In addition, the interferon pathways inhibited by PRRSV infection were reversed by the inhibition of glycolysis. These findings may facilitate further investigation of the role of glycolysis in PRRSV replication.
Porcine respiratory and reproductive syndrome virus/physiology* ; Glycolysis ; Swine ; Animals ; Virus Replication ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics* ; Macrophages, Alveolar/metabolism* ; Porcine Reproductive and Respiratory Syndrome/virology* ; Cells, Cultured ; RNA, Small Interfering/genetics*

OBJECTIVETo determine the physical and magnetic properties of superparamagnetic iron oxide (SPIO) nanoparticle prepared in our laboratory and evaluate its possibility for use as contrast agents in magnetic resonance imaging (MRI).

METHODSThe SPIO nanoparticle was obtained by means of classical coprecipitation in dextran solution and its size determined by electron microscopy and photon-correlation spectroscopy. The iron content was determined by phenanthroline photometry, and T(2) values as well as relaxivity evaluated with a clinical MR system at 1.5T.

RESULTSDextran-coated magnetite particles with a hydrodynamic diameter of 85.9 nm were prepared. The iron core size was 15 nm and the formation of Fe(3)O(4) crystal in SPIO nanoparticles was confirmed by X-ray diffraction (XRD) analysis. These particles possessed some characteristics of superparamagnetic and show a smaller spin-spin relaxation, with relaxivity and saturation magnetization of 0.1567 mmol(-1)/ms(-1) and 80 emu/g Fe, respectively.

CONCLUSIONSA stable SPIO nanoparticle with a dextran coating have been developed, and in vitro evaluation of its physical and magnetic properties suggests its potential for use as the contrast agent in MRI.

Humans ; Iron ; chemistry ; Magnetic Resonance Imaging ; methods ; Nanoparticles ; chemistry ; Oxides ; chemical synthesis ; chemistry ; X-Ray Diffraction