ObjectiveTo investigate the effects of targeted silencing of Runt-related Transcription Factor 3 （RUNX3） on the proliferation and migration of Mouse Hepatic Stellate Cells （HSCs）， as well as subsequent collagen deposition. MethodsMouse hepatic stellate cell line （JS-1） was selected and then morphologically observed and identified under a microscope. After the cells had fully adhered， they were treated with 5 ng/mL of transforming growth factor beta 1 （TGF-β₁） for 24 hours to induce hepatic stellate cell activation. Furthermore， a RUNX3 silencing model was established using RUNX3 lentiviral infection. The experiment was divided into four groups： Control group， TGF-β₁ group， TGF-β₁+siRNA-NC group， and TGF-β₁+siRNA-RUNX3 group. Protein expression changes of RUNX3， alpha-smooth muscle actin （α-SMA）， and Alpha 1 type I collagen （Collagen I） were detected using Western blot method. Cellular immunofluorescence assays were employed to investigate the deposition changes of α-SMA and RUNX3 in hepatic stellate cells. RT-qPCR was utilized to examine the mRNA expression changes of RUNX3， α-SMA， and Collagen I. The proliferative capacity of hepatic stellate cells was assessed using Edu staining. The migratory ability of hepatic stellate cells was evaluated through wound healing assays and Transwell migration experiments. ResultsCompared with Control group， a significant elevation in RUNX3 was observed in the TGF-β₁-induced activated HSCs （P<0.01）. Meanwhile， the protein and mRNA levels of fibrosis-related markers and α-SMA and Collagen I were significantly upregulated （P<0.001）. Additionally， the proliferation and migration capabilities of HSCs were significantly enhanced （P<0.001）. In contrast， when compared to TGF-β₁+siRNA-NC group， TGF-β₁+siRNA-RUNX3 group exhibited a notable decrease in RUNX3 and other related indicators， such as the protein and mRNA levels of α-SMA and Collagen I （P<0.05）. Concurrently， the proliferation and migration capabilities of HSCs were significantly inhibited in TGF-β₁+siRNA-RUNX3 group （P<0.01）. ConclusionSilencing RUNX3 can inhibit the deposition of collagen and the proliferation and migration of hepatic stellate cells. Conversely， RUNX3 promotes the proliferation and migration capabilities of HSCs， thereby facilitating the activation of HSC.

This study aims to explore the mechanism of Huanglian Jiedu Decoction(HJD) in treating acute liver injury(ALI) in the mouse model of sepsis induced by lipopolysaccharide(LPS). Fifty-four male C57BL/6 mice were randomized into six groups: blank group, model group, low-, medium-, and high-dose group HJD, and dexamethasone group. The mouse model of sepsis was established by intraperitoneal injection of LPS after 7 days of gavage with HJD, and dexamethasone(0.2 mL) was injected intraperitoneally 1.5 h after modeling. The murine sepsis score(MSS) was recorded 12 h after modeling. The levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the liver tissue and tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in the serum were measured by ELISA. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of the mouse liver. The content of light chain 3 of microtubule-associated protein 1(LC3) was detected by immunofluorescence, and that of sirtuin 1(SIRT1) was detected by immunohistochemistry. The mRNA levels of adenosine 5'-monophosphate-activated protein kinase(AMPK), LC3, and P62 were detected by RT-PCR. Western blot was employed to determine the protein levels of AMPK, p-AMPK, and SIRT1 in the liver tissue. The results showed that compared with model group, drug interventions decreased the MSS and liver injury indicators, lowered the levels of inflammatory cytokines, improved the liver tissue structure, upregulated the protein levels of of p-AMPK/AMPK and SIRT1 and the mRNA levels of AMPK and LC3, and downregulated the mRNA level of P62. To sum up, HJD can regulate the autophagy level and reduce inflammation to ameliorate acute liver injury in septic mice by activating the AMPK/SIRT1 autophagy pathway.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Sirtuin 1/genetics* ; Male ; Mice ; Sepsis/metabolism* ; Mice, Inbred C57BL ; Autophagy/drug effects* ; AMP-Activated Protein Kinases/genetics* ; Liver/metabolism* ; Humans ; Signal Transduction/drug effects* ; Disease Models, Animal ; Tumor Necrosis Factor-alpha/genetics*

The auxin/indole-3-acetic acid (Aux/IAA) gene family is an important regulator for plant growth hormone signaling, involved in plant growth, development, as well as response to environmental stresses. In the present study, we identified SmIAA7 which is potentially associated with Salvia miltiorrhiza leaf development through comparatively analyzed the transcriptome data from different pinnate leaves. SmIAA7 was successfully isolated from S. miltiorrhiza using the specific primers. Then subsequent bioinformatic analysis, prokaryotic expression and purification, subcellular localization, and induction expression analysis under auxin and abiotic stress were performed. The full-length of SmIAA7 contained an ORF of 684 bp encoding a protein of 227 amino acid with a molecular weight of 25.3 kD. Conserved domain analysis showed that SmIAA7 contains the conserved Aux_IAA domain (pfam02309). Sequence analysis and phylogenetic tree analysis results indicated SmIAA7 was phylogenetically close to IAA7 and IAA14 from other plants, suggesting SmIAA7 involved in plant growth and development as well as response to environmental stresses. The prokaryotic expression vector pET28a-SmIAA7 was constructed and SmIAA7 recombinant protein was successfully expressed in E. coli Rosetta (DE3) strain. Subcellular localization experiment demonstrated that SmIAA7 localized in the nucleus of plant cells. Real-time fluorescence quantitative PCR results showed that the expression level of SmIAA7 was upregulated in response to auxin. Drought, low temperature, and salt stress significantly increased the transcript level of SmIAA7 gene. This study lays a foundation for further elucidating the role of SmIAA7 in leaf development, signal transduction, and stress defense in S. miltiorrhiza.

Aim To explore the influence of demethy-lase fat mass and obesity-related genes(FTO)on the abnormal contractile function of diabetic coronary smooth muscle.Methods Smooth muscle specific FTO knockout mice(FTOSMKO)were prepared by Cre-loxP recombinant technology.They were divided into four groups:control(WT)group,diabetes model group(DM),FTO knockout group(FTOSMKO),and FTOSMKO diabetic group(FTOSMKO-DM),with 15 mice in each group.Diabetic mice were prepared by intraperitoneal injection of streptozotocin(STZ);the remaining mice were injected with an equal amount of citric acid-sodi-um citrate buffer.The effects of 5-HTon the contractile response of coronary artery smooth muscle in the four groups of mice were observed by the technique of small-vessel ring tensiometry.Western blot and Dot blot were used to detect the changes of FTO protein and N6-methyladenine(m6A)methylation modification levels in mouse vascular tissues.Results Compared with the WT group,the DM group had significantly higher blood glucose(P＜0.01)and lower body weight(P＜0.05);the level of FTO protein in aorta of DM group increased(P＜0.01),and the level of m6A methylation modification decreased(P＜0.01).The 5-HT-induced contractile response significantly de-creased in the DM group compared with the WT group(P＜0.01),whereas the contractile response signifi-cantly increased in the FTOSMKO-DM group compared with the DM group(P＜0.01);the non-L-type calci-um channel-mediated vascular smooth muscle contrac-tile response was enhanced in the FTOSMKO-DM group,among which,the contractions induced by 1,4,5-triphosphate inositol receptor(IP3R)and caffeine-ac-tivated ranine receptor(RyR)mediated by sarcoplas-mic reticulum calcium release both significantly in-creased(P＜0.05).Conclusions Specific knock-down of smooth muscle FTO improves coronary artery responsiveness to the vasoconstrictor 5-HT in diabetic mice,which may be related to abnormalities in the FTO-mediated 5-HT receptor signaling pathway.

Objective Network pharmacology and molecular docking techniques were used to predict the potential mechanisms by which the active components of Piper nigrum(PN)regulate depressive-like behaviors in chronic restraint stress(CRS)mice.Methods The major chemical components and targets of PN were screened using the Traditional Chinese Medicine Systems Pharmacology database.Targets related to ferroptosis and depression were obtained from the Online Mendelian Inheritance in Man,GeneCards,and FerrDB databases.The intersecting targets were then subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Gnomes(KEGG)pathway enrichment analyses,and molecular docking was performed to validate the binding capacities between the core targets and their corresponding active components.Finally,we established a CRS mouse model.Mice were treated with PN 75,150,and 300 mg/kg for 4 weeks,followed by behavioral assessments and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)to verify the expression of core genes.Results Nine active components were screened from PN,corresponding to 27 targets,and 8377 targets related to depression and 547 targets associated with ferroptosis were screened from the databases.The intersection of these three sets resulted in 25 target genes.KEGG enrichment analysis revealed that these core targets were predominantly enriched in signaling pathways,including cholinergic synapses,serotonergic synapses,and neuroactive ligand-receptor interactions.Molecular docking result showed that the main active components of PN had strong binding affinities for the targets CHRM2,SLC6A4,PTGS2,and SLC6A2.Behavioral assessments demonstrated that PN significantly increased the sucrose preference index(P＜0.01,P＜0.001),reduced immobility time in the tail suspension and forced swimming tests(P＜0.01,P＜0.001),and enhanced exploratory behavior in the open field test(P＜0.05.P＜0.01,P＜0.001).PN significantly reduced the serum levels of inflammation markers(P＜0.05.P＜0.01,P＜0.001),as shown by enzyme-linked immunosorbent assay,and neurotransmitter analysis revealed that PN significantly increased the levels of serotonin and acetylcholine in the mouse hippocampus(P＜0.05).RT-qPCR showed that PN demonstrated the mRNA expression of SLC6A4(P＜0.05.P＜0.01,P＜0.001).Conclusions PN may improve depressive-like behavior in mice by modulating serotonin and acetylcholine levels,inhibiting inflammatory responses,participating in immune regulation,and exerting neuroprotective effects.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective:To investigate the effect of traditional Chinese medicine chronic disease management model based on empowerment theory in patients with chronic heart failure(CHF).Methods:A total of 115 CHF patients admitted in Guangdong Provincial Hospital of Chinese Medicine between January 2020 and December 2021 were se-lected.Patients received traditional Chinese medicine chronic disease management model based on empowerment theory according to voluntary principle,and were followed up for 12 months.Exercise capacity,scores of Tradition-al Chinese Medicine Symptom Grading and Quantification Scale,Hospital Anxiety and Depression Scale(HADS)and Minnesota Living with Heart Failure Questionnaire(MLHFQ)were compared between before and after inter-vention.Results:Compared to before intervention,scores of Traditional Chinese Medicine Symptom Grading and Quantification Scale[(6.40±6.11)points vs.(8.88±6.72)points],HADS[(5.95±4.68)points vs.(7.69±5.95)points],MLHFQ[(13.10±10.54)points vs.(25.53±11.16)points]and 3m round-trip movement time[(7.54±1.70)s vs.(8.86±3.65)s]were significantly lower,and right hand grip strength[(27.23±10.49)kg vs.(26.10±9.94)kg]and 6-minute walking distance[(464.79±80.78)m vs.(415.55±79.33)m]were sig-nificantly higher after 12-month intervention(P＜0.05 or＜0.01).Conclusion:The traditional Chinese medicine chronic disease management model based on empowerment theory may improve clinical symptoms of traditional Chi-nese medicine,mental state,exercise capacity and quality of life in patients with chronic heart failure.

Objective:This study aimed to evaluate whether intra-articular delivery of conditioned medium(CM)derived from stem cells of human exfoliated deciduous teeth(SHED)could influence the progression of condylar cartilage degeneration in a rat model of temporomandibular joint osteoarthritis(TMJ OA).Methods:Sixty 8-week-old male SD rats were randomly divided into control group(CON group),intraarticular injection of MIA induced TMJ OA model group(MIA group),and injection of SHED condi-tioned medium 1 week after MIA modeling for treatment group(SHED-CM group),with 20 animals in each group.Histological sec-tions,HE,Safranine O-solid green staining,Col Ⅱ immunohistochemical staining,and TUNEL staining were performed 2 and 4 weeks after the start of treatment.Western blotting and RT-qPCR were used to detect the key molecules of apoptosis,cleaved-CASP3,BAX and BCL2,pro-inflammatory related factors IL-1β,IL-6,TNFα,MMP3,ADAMTS5,and the MAPK pathway-related molecules p-ERK,ERK,p-P38 and P38.Results:Compared with the CON group and SHED-CM group,the condyle chondrocytes in the MIA group had disordered arrange-ment,interrupted layers,significantly thickened fibrous layers(P＜0.001),and significantly increased Mankin's OA histological score(P＜0.001).In the MIA group,both the Safranin O-positive area ratio and the proportion of ColⅡ-positive regions were markedly reduced compared with the CON and SHED-CM groups(P＜0.001).Conversely,the proportion of TUNEL-positive cells was substantially higher than in the other two groups(both P＜0.001).Western blot analysis further demonstrated that apoptotic markers(cleaved-CASP3,BAX/BCL2)and MAPK pathway-related proteins(p-ERK,ERK,p-P38,P38)were expressed at significantly elevated levels in the MIA group relative to CON and SHED-CM groups(BAX/BCL2:P＜0.05;cleaved-CASP3:P＜0.01;p-P38/P38:P＜0.001;p-ERK/ERK:P＜0.01).Similarly,qRT-PCR revealed upregulated expression of inflammatory mediators,including IL-1 β(P＜0.001),IL-6(P＜0.01),TNFα(P＜0.01),MMP3(P＜0.001),and ADAMTS5(P＜0.05),in the MIA group compared with the CON and SHED-CM groups.Conclusion:SHED-CM treatment can ef-fectively reverse MIA-induced condylar cartilage degeneration of TMJ OA in rats.

Objective To investigate clinical therapeutic value of combined posterior nasal nerve and anterior ethmoidal nerve ablation in patients with nasal septal deviation and moderate to severe persistent allergic rhinitis.Methods A total of 47 patients diagnosed with nasal septal deviation and moderate to severe persistent allergic rhinitis from April to December 2024 were divided into two groups.The control group(n=22)underwent septoplasty and bilateral inferior turbinate out-fracture,with postoperative symptom control managed by budesonide nasal spray.The experimental group(n=25)received the same septoplasty and turbinate surgery,supplemented by bilateral posterior nasal nerve and anterior ethmoidal nerve ablation.Symptom improvement was compared between groups by using the Visual Analogue Scale(VAS)and Rhinoconjunctivitis Quality of Life Questionnaire(RQLQ)scores preoperatively and3 months postoperatively.Results The operation time of the control group was(36.3±5.9)min,significantly shorter than that of the experimental group[(59.4±6.6)min,t=12.496,P=0.000].Both groups of patients showed significant improvement in VAS and PQLQ scores for nasal symptoms such as nasal congestion,runny nose,sneezing,and itching at 3 months after surgery compared to preoperative levels(all P=0.000).The mean VAS score of the four symptoms in the control group was(4.6±0.9)points at3 months after surgery,which was significantly lower than that before surgery[(6.3±1.1)points,t=9.796,P=0.000].The mean VAS score of the four symptoms in the experimental group was1.0(0.3-4.3)points at3 months after surgery,which was significantly lower than that before surgery[7.0(4.5-9.0)points,Z=-4.376,P=0.000].The improvement rate of the VAS score in the experimental group was(82.4±14.2)%,significantly higher than that in the control group[(26.9±11.7)%,t=14.510,P=0.000].At3 months after surgery,the RQLQ score of the control group[(2.3±0.8)points]was significantly lower than that before surgery[(3.3±0.8)points,t=10.055,P=0.000].The RQLQ score of the experimental group after surgery was 1.4(0.8-3.5)points,which was significantly lower than the preoperative score[3.6(1.5-6.1)points,Z=-4.373,P=0.000].The improvement rate of RQLQ score in the experimental group was(53.0±14.6)%,significantly higher than that in the control group[(30.2±13.4)%,t=5.555,P=0.000].Conclusion Postnasal nerve and anterior ethmoidal nerve ablation combined with nasal septal deviation correction can significantly improve nasal symptoms and quality of life in patients with nasal septal deviation complicated with moderate to severe allergic rhinitis compared with simple nasal septal deviation correction,but the operation time is prolonged.