Objective To explore the mechanism of honey-processed Hedysari Radix in the regulation of intestinal immunity in rats with spleen qi deficiency,which was based on G protein-coupled receptor 41(GPR41)/GPR43-mediated mitogen-activated protein kinase(MAPK)signaling pathway.Methods The three-factor composite modeling method of eating disorder,diarrhea and fatigue was used to establish a model of spleen qi deficiency,and the rats were randomly divided into model,honey-processed Hedysari Radix,probiotics and blank groups with 15 rats per group.The honey-processed Hedysari Radix group was given by gavage 12.6 g·kg-1 aqueous extract of honey-processed Hedysari Radix.The probiotics group was given 0.625 g·kg-1 bifidobacterium triple viable solution by gavage.The blank and model groups were given the same dose of distilled water by gavage.Four groups were treated for 15 d with once a day.The expression levels of GPR41,GPR43,P38 MAPK,c-Jun N-terminal kinase(JNK)and extracellular regulatory protein kinase 1/2(ERK1/2)in colon tissues were detected by Western blotting.Results The relative expression levels of GPR41 in the blank,model,honey-processed Hedysari Radix and probiotics groups were 0.95±0.07,0.45±0.03,0.84±0.19 and 0.86±0.20;the relative expression levels of GPR43 were 1.17±0.11,0.41±0.06,0.66±0.03 and 0.57±0.01;the phosphorylated ERK1/2/ERK1/2 ratios were 0.16±0.01,0.43±0.01,0.39±0.01 and 0.36±0.02;the phosphorylated JNK/JNK ratios were 0.58±0.05,1.47±0.10,0.90±0.11 and 0.90±0.11;the phosphorylated P38 MAPK/P38 MAPK ratios were 1.77±0.33,3.19±0.03,2.01±0.17 and 2.23±0.59,respectively.Compared with the model group,the differences of above indexes were statistically significant in the honey-processed Hedysari Radix and probiotics groups(P＜0.05,P＜0.01).Conclusion The mechanism of honey-processed Hedysari Radix regulating intestinal immunity in rats with spleen qi deficiency is related to the regulation of GPR41/GPR43 mediated MAPK signaling pathway.

Objective:To explore the internal mechanism by which self-evaluation influences non-suicidal self-injury (NSSI) in adolescent patients with depressive disorder.Methods:Clinical data from 214 adolescent patients with depressive disorder hospitalized at Suzhou Guangji Hospital from March 2022 to January 2024 were prospectively collected. According to the DSM-5 diagnostic criteria for NSSI, participants were divided into an NSSI group (158 cases [38 males, 120 females, age 12-17 (14.2±1.5) years]) and a non-NSSI group (56 cases [20 males, 36 females, age 12-18 (14.5±1.8) years]). A self-developed basic information questionnaire was used to collect demographic data. Standardized tools including the Self-rating Depression Scale (SDS), Inventory of Parent and Peer Attachment (IPPA), Self-acceptance Questionnaire (SAQ), and Perceived Stress Scale-10 (PSS-10) were used to assess their depression level, parental attachment level, self-evaluation/acceptance level, perceived stress level, and other relevant psychological characteristics. Differences in psychological characteristics between the two groups were compared. Logistic regression, correlation analysis, and mediation effect models were used to explore the relationships between variables and NSSI and their mechanisms.Results:The NSSI group had significantly higher total scores on the SDS (41.3±7.7 vs. 34.4±9.3) and PSS-10 (25.5±6.1 vs. 21.3±6.5) than the non-NSSI group ( F=29.12, F=18.17, respectively; all P<0.001). Conversely, the NSSI group had significantly lower total scores on the SAQ (31.2±8.8 vs. 35.9±8.9) and IPPA (56.3±13.6 vs. 63.4±13.8) compared to the non-NSSI group ( F=11.24, F=10.84, respectively; all P<0.001). Stepwise logistic regression analysis identified depression level (SDS total score, OR=1.12, 95% CI: 1.05-1.19), self-evaluation (SAQ subscale score, OR=1.17, 95% CI: 1.04-1.31), and perceived stress (PSS-10 total score, OR=1.11, 95% CI: 1.01-1.22) as predictors of NSSI (all P<0.05). Chain mediation analysis showed that self-evaluation had a significant positive direct effect ( β=0.025, P<0.01) and a negative indirect effect ( β=-0.038, P<0.001) on NSSI, with a negative total effect ( β=-0.012, P<0.05). The indirect effect was realized through three pathways: a single mediation pathway of self-evaluation via perceived stress ( β=-0.016), a single mediation pathway of self-evaluation via depression ( β=-0.011), and a chain mediation pathway of self-evaluation via perceived stress and depression ( β=-0.011) (all P<0.05). Conclusion:Self-evaluation influences NSSI behavior through a dual mechanism involving both direct and indirect effects. The indirect protective effect is primarily achieved by reducing perceived stress and depression levels.

Objective:To investigate therapeutic effect and mechanism of sanguinarine on postherpetic neuralgia(PHN)rats by modulating C-X-C chemokine ligand 12(CXCL12)/C-X-C chemokine receptor 4(CXCR4)signaling pathway.Methods:SD rats were randomly grouped into control group,model group,low-dose(50 mg/kg)sanguinarine group,high-dose(100 mg/kg)sanguina-rine group,NUCC-390(CXCL12/CXCR4 signal activator,2.2 mg/kg)group,high-dose(100 mg/kg)sanguinarine+NUCC-390(2.2 mg/kg)group,with 10 rats in each group.Rats in model group and drug-treated groups were injected with resin toxin(RTX)by intraperitoneal injection to induce PHN model,rats in control group were intraperitoneally injected with an equal dose of normal saline containing 10%Tween 80 and 10%ethanol.After treatment of sanguinarine and NUCC-390,symptoms of long-term spontaneous pain,mechanical hyperalgesia and thermal hyperalgesia were detected,number of spontaneous paw withdrawal reflexes,paw with-drawal threshold to mechanical stimulation(PWMT),and response latency to thermal stimulation(PWTL)were compared;spinal cord nerve cell apoptosis was detected by TUNEL staining;ELISA was used to detect levels of inflammatory factors TNF-α,IL-1β,cyclooxygenase-2(COX-2)in rat spinal cord tissue and serum;Western blot was used to detect expressions of CXCL12/CXCR4 path-way-related proteins in spinal cord tissues of rats in each group.Results:Compared with control group,PWMT of model group was obviously decreased(P<0.05),number of spontaneous foot withdrawal reflexes,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were obviously increased(P<0.05).Compared with model group,PWMT of rats in low-dose sanguinarine group and high-dose sanguinarine group was increased(P<0.05),number of spontaneous foot withdrawal reflexes,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were all decreased(P<0.05);PWMT of rats in NUCC-390 group was decreased(P<0.05),number of spontaneous foot withdrawal reflex,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were increased(P<0.05).Compared with high-dose sanguinarine group,PWMT of rats in high-dose sanguinarine+NUCC-390 group was decreased(P<0.05),number of spontaneous foot withdrawal reflex,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were increased(P<0.05).Conclusion:Sanguinarine can reduce expression of inflammatory factors by down-regulating CXCL12/CXCR4 signaling pathway,thereby preventing occurrence of inflammatory response in PHN rats,inhibiting apoptosis of spinal nerve cells,and finally reducing long-term spontaneous pain,mechanical allodynia and thermal hypoalgesia in rats.

Aim To investigate the effect of chrysoeriol on pulmonary vascular remodeling in pulmonary hyper-tension by animal experiments combined with cell ex-periments,and to explore its potential therapeutic tar-gets by network pharmacology.Methods The target of chrysoeriol was collected in Targetnet,SEA and SwissTargetPrediction database.Pulmonary arterial hy-pertension(PAH)targets were collected in the Dis-GeNET and GeneCards databases,and PPI network map was drawn in the STRING database,and key tar-gets were screened.The GO and KEGG pathway en-richment analysis was carried out through DAVID data-base and Weishengxing platform.AutoDock software was used for molecular docking of key core targets.The PAH model of rats was constructed,and the pulmo-nary hemodynamics and vascular remodeling were de-tected by echocardiography,HE and Masson staining.Primary pulmonary smooth muscle cells were extracted,and the effects of drugs on pathway proteins were de-tected in vitro.Results The results of network phar-macology showed that chrysoeriol exerted therapeutic effects on pulmonary hypertension by affecting key tar-gets such as AKT1,SRC,EGFR,MMP9 and gsk3 β,and signaling pathways such as EGFR and PI3K-AKT.Molecular docking showed that chrysoeriol had good binding ability with 5 key target genes.Animal experi-ments showed that the pulmonary hemodynamic func-tion of PAH rats was significantly improved after ad-ministration of chrysoeriol.The remodeling of small pulmonary arteries was significantly reduced.Cell ex-periments showed that chrysoeriol could inhibit the ex-pression of proliferation,migration and phenotypic transformation genes.Conclusion Chrysoeriol may play a role in the treatment of pulmonary hypertension through multiple targets.

Objective To investigate the mechanism by which suppressor of cytokine signaling 3(SOCS3)regulates microglial inflammation through nuclear factor-kappaB(NF-κB),providing novel mechanistic insights into microglial involvement in Parkinson's disease(PD)pathogenesis.Methods ① Ten male C57BL/6 mice(12 weeks old,weighing 20～25 g)were subjected to intraperitoneal injection of 15 mg/kg MPTP to establish a PD model.Rotarod test was used to assess motor function.Western blotting was employed to detect the protein expression of tyrosine hydroxylase(TH)and ionized calcium-binding adapter molecule 1(IBA-1)in the substantia nigra.RT-qPCR was utilized to measure the mRNA level of SOCS3 in the substantia nigra.Immunohistochemistry was performed to assess NF-κB p65 subunit expression.The expression of SOCS3,NF-κB and p-NF-κB was measured with Western blotting.② Microglial cell line BV2 was stimulated with 1 000 ng/mL lipopolysaccharide(LPS)for 6 h to establish an inflammatory model.Subsequently,SOCS3 was knocked down.NF-κB inhibitor BAY 11-7082 was used to treat the cells.RT-qPCR and Western blotting were used to measure the expression of SOCS3 at mRNA and protein levels.Western blotting was also applied to detect the expression of NF-κB and p-NF-κB,and ELISA was conducted to measure TNF-α and IL-1β levels in the culture supernatant.Immunofluorescence assay was carried out to localize NF-κB(nuclear vs cytoplasmic).③ A co-culture system of BV2 microglia and N2a neuroblastoma cells was established to investigate the regulatory effects of microglia on neuronal cells.MTT assay and TUNEL staining were used respectively to determine cell viability and apoptosis of N2a cells.Results ① Compared to the control mice,the PD mouse model exhibited reduced rotarod fall latency,down-regulation in TH and SOCS3(P<0.01),up-regulation in IBA-1 and increased p-NF-κB/NF-κB ratio(P<0.01).② In BV2 cells,LPS stimulation increased TNF-α,IL-1β,and p-NF-κB/NF-κB ratio(P<0.01),while down-regulated SOCS3 expression(P<0.01).SOCS3 knockdown in LPS-stimulated BV2 cells further increased the p-NF-κB/NF-κB ratio(P<0.01),increased nuclear localization of NF-κB,and elevated TNF-α and IL-1β levels(P<0.01).BAY 11-7082 treatment in these SOCS3-knockdown,LPS-stimulated cells resulted in reduced p-NF-κB/NF-κB ratio,TNF-α,and IL-1β(P<0.01),and decreased NF-κB nuclear distribution.③ LPS-stimulated BV2 cells reduced cell viability and increased cell apoptosis in N2a cells(P<0.01).SOCS3 knockdown in BV2 cells exacerbated the reduction in N2a cell viability(P<0.01)and the increase in cell apoptosis in N2a cells(P<0.01).BAY 11-7082 treatment of these SOCS3-knockdown BV2 microglia attenuated the reduction in N2a cell viability and decreased apoptosis in N2a cells(P<0.01).Conclusion SOCS3 inhibits microglia inflammatory response through down-regulation of NF-kB activity,and in turn attenuates neuronal cell death and ameliorates PD nerve injury.

Objective To demonstrate that neferine(Nef)alleviates Parkinson's disease(PD)by inhibiting microglial pyroptosis mediated through the reactive oxygen species(ROS)/NOD-like receptor protein 3(NLRP3)/Caspase-1 pathway.Methods BV2 microglial cells were divided into:control group,lipopolysaccharides(LPS)-adenosine triphosphate(ATP)group,and LPS-ATP+Nef group.Pyroptosis was induced by 1 μg/mL LPS+5 mmol/L ATP,with 2 mmol/L Nef pretreatment.Eighteen 10-12-week-old male C57BL/6 mice(22～25 g)were randomly assigned to:control(n=6),1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)(n=6),and MPTP+Nef(n=6)groups.Detection methods included:flow cytometry for pyroptosis,Cell Counting Kit-8(CCK-8)for viability,2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)for ROS,commercial kits for malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),ELISA/Western blot for interleukin-1β(IL-1β)/IL-18,immunofluorescence/immunohistochemistry for NLRP3/Caspase-1,tyrosine hydroxylase(TH)immunohistochemistry,hematoxylin-eosin staining for neuropathology,and modified neurological severity score(mNSS).Results Versus control,LPS-ATP group showed decreased viability(P=0.002),increased pyroptosis(P<0.001),elevated ROS(P<0.001)/MDA(P<0.001)/IL-1β(P<0.001)/IL-18(P<0.001),upregulated NLRP3(P<0.001)/Caspase-1(P<0.001),and reduced GSH(P<0.001)/SOD(P<0.001).Nef treatment reversed these effects(all P<0.05).According to the results of murine studies,compared with the control group,the MPTP group had increased mNSS(P<0.001)/tissue ROS(P<0.001),downregulated TH(P<0.001),upregulated NLRP3(P<0.001)/Caspase-1(P<0.001).Nef treatment significantly attenuated the MPTP-induced deleterious effects(P<0.05).Histopathological analysis revealed that control group exhibited uniformly distributed hippocampal neurons with distinct nuclear morphology;MPTP group showed neuronal swelling,interstitial edema,and nuclear atrophy;MPTP+Nef group demonstrated ameliorated neuronal damage.Conclusion Nef inhibits microglial pyroptosis via ROS/NLRP3/Caspase-1 axis,ameliorating PD neuroinflammation and pathology.

Aim To investigate the effect of chrysoeriol on pulmonary vascular remodeling in pulmonary hyper-tension by animal experiments combined with cell ex-periments,and to explore its potential therapeutic tar-gets by network pharmacology.Methods The target of chrysoeriol was collected in Targetnet,SEA and SwissTargetPrediction database.Pulmonary arterial hy-pertension(PAH)targets were collected in the Dis-GeNET and GeneCards databases,and PPI network map was drawn in the STRING database,and key tar-gets were screened.The GO and KEGG pathway en-richment analysis was carried out through DAVID data-base and Weishengxing platform.AutoDock software was used for molecular docking of key core targets.The PAH model of rats was constructed,and the pulmo-nary hemodynamics and vascular remodeling were de-tected by echocardiography,HE and Masson staining.Primary pulmonary smooth muscle cells were extracted,and the effects of drugs on pathway proteins were de-tected in vitro.Results The results of network phar-macology showed that chrysoeriol exerted therapeutic effects on pulmonary hypertension by affecting key tar-gets such as AKT1,SRC,EGFR,MMP9 and gsk3 β,and signaling pathways such as EGFR and PI3K-AKT.Molecular docking showed that chrysoeriol had good binding ability with 5 key target genes.Animal experi-ments showed that the pulmonary hemodynamic func-tion of PAH rats was significantly improved after ad-ministration of chrysoeriol.The remodeling of small pulmonary arteries was significantly reduced.Cell ex-periments showed that chrysoeriol could inhibit the ex-pression of proliferation,migration and phenotypic transformation genes.Conclusion Chrysoeriol may play a role in the treatment of pulmonary hypertension through multiple targets.

Objective To explore the mechanism of honey-processed Hedysari Radix in the regulation of intestinal immunity in rats with spleen qi deficiency,which was based on G protein-coupled receptor 41(GPR41)/GPR43-mediated mitogen-activated protein kinase(MAPK)signaling pathway.Methods The three-factor composite modeling method of eating disorder,diarrhea and fatigue was used to establish a model of spleen qi deficiency,and the rats were randomly divided into model,honey-processed Hedysari Radix,probiotics and blank groups with 15 rats per group.The honey-processed Hedysari Radix group was given by gavage 12.6 g·kg-1 aqueous extract of honey-processed Hedysari Radix.The probiotics group was given 0.625 g·kg-1 bifidobacterium triple viable solution by gavage.The blank and model groups were given the same dose of distilled water by gavage.Four groups were treated for 15 d with once a day.The expression levels of GPR41,GPR43,P38 MAPK,c-Jun N-terminal kinase(JNK)and extracellular regulatory protein kinase 1/2(ERK1/2)in colon tissues were detected by Western blotting.Results The relative expression levels of GPR41 in the blank,model,honey-processed Hedysari Radix and probiotics groups were 0.95±0.07,0.45±0.03,0.84±0.19 and 0.86±0.20;the relative expression levels of GPR43 were 1.17±0.11,0.41±0.06,0.66±0.03 and 0.57±0.01;the phosphorylated ERK1/2/ERK1/2 ratios were 0.16±0.01,0.43±0.01,0.39±0.01 and 0.36±0.02;the phosphorylated JNK/JNK ratios were 0.58±0.05,1.47±0.10,0.90±0.11 and 0.90±0.11;the phosphorylated P38 MAPK/P38 MAPK ratios were 1.77±0.33,3.19±0.03,2.01±0.17 and 2.23±0.59,respectively.Compared with the model group,the differences of above indexes were statistically significant in the honey-processed Hedysari Radix and probiotics groups(P＜0.05,P＜0.01).Conclusion The mechanism of honey-processed Hedysari Radix regulating intestinal immunity in rats with spleen qi deficiency is related to the regulation of GPR41/GPR43 mediated MAPK signaling pathway.

Objective:To investigate therapeutic effect and mechanism of sanguinarine on postherpetic neuralgia(PHN)rats by modulating C-X-C chemokine ligand 12(CXCL12)/C-X-C chemokine receptor 4(CXCR4)signaling pathway.Methods:SD rats were randomly grouped into control group,model group,low-dose(50 mg/kg)sanguinarine group,high-dose(100 mg/kg)sanguina-rine group,NUCC-390(CXCL12/CXCR4 signal activator,2.2 mg/kg)group,high-dose(100 mg/kg)sanguinarine+NUCC-390(2.2 mg/kg)group,with 10 rats in each group.Rats in model group and drug-treated groups were injected with resin toxin(RTX)by intraperitoneal injection to induce PHN model,rats in control group were intraperitoneally injected with an equal dose of normal saline containing 10%Tween 80 and 10%ethanol.After treatment of sanguinarine and NUCC-390,symptoms of long-term spontaneous pain,mechanical hyperalgesia and thermal hyperalgesia were detected,number of spontaneous paw withdrawal reflexes,paw with-drawal threshold to mechanical stimulation(PWMT),and response latency to thermal stimulation(PWTL)were compared;spinal cord nerve cell apoptosis was detected by TUNEL staining;ELISA was used to detect levels of inflammatory factors TNF-α,IL-1β,cyclooxygenase-2(COX-2)in rat spinal cord tissue and serum;Western blot was used to detect expressions of CXCL12/CXCR4 path-way-related proteins in spinal cord tissues of rats in each group.Results:Compared with control group,PWMT of model group was obviously decreased(P<0.05),number of spontaneous foot withdrawal reflexes,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were obviously increased(P<0.05).Compared with model group,PWMT of rats in low-dose sanguinarine group and high-dose sanguinarine group was increased(P<0.05),number of spontaneous foot withdrawal reflexes,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were all decreased(P<0.05);PWMT of rats in NUCC-390 group was decreased(P<0.05),number of spontaneous foot withdrawal reflex,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were increased(P<0.05).Compared with high-dose sanguinarine group,PWMT of rats in high-dose sanguinarine+NUCC-390 group was decreased(P<0.05),number of spontaneous foot withdrawal reflex,PWTL,spinal nerve cell apoptosis index,levels of TNF-α,IL-1β,COX-2 in spinal cord tissue and serum,and protein expressions of CXCL12 and CXCR4 in spinal cord tissue were increased(P<0.05).Conclusion:Sanguinarine can reduce expression of inflammatory factors by down-regulating CXCL12/CXCR4 signaling pathway,thereby preventing occurrence of inflammatory response in PHN rats,inhibiting apoptosis of spinal nerve cells,and finally reducing long-term spontaneous pain,mechanical allodynia and thermal hypoalgesia in rats.