Background: Malignant pleural effusion (MPE), a common complication of advanced cancers, is associated with poor prognosis and reduced quality of life. Although host–tumor interactions are known to drive MPE development, the associated immune dynamics during disease progression remain unclear. Using a Lewis lung carcinoma-induced MPE model in C57BL/6JNidfc mice, we systematically evaluated general parameters and immune cell changes at two-day intervals throughout disease progression. Results: The day of Lewis lung carcinoma cell injection into the pleural space was designated as day 0. By day 10 post-injection (p.i.), MPE-bearing mice exhibited ~ 10% body weight loss, marking the experimental endpoint. Pleural tumor mass and pleural effusion volume were minimal up to day 4 p.i. but increased sharply from day 6 onward.CD45⁺ immune cell counts rose over time, and days 6, 8, and 10 p.i. marked key stages of MPE progression. On day 6, B cells, T cells, and natural killer cells, but not macrophages and neutrophils, increased significantly compared to earlier timepoints. By day 8, all immune cell subsets except T cells exceeded day 6 levels, and at day 10, natural killer cell numbers declined while others continued to increase. Besides, the numbers of CD8⁺ T cells, Th1 cells, regulatory T cells, and M2 macrophages progressively increased from day 6 to 10. Based on these data, days 6 and 10 were defined as early and advanced MPE stages, respectively, with distinct immune phenotypes. In advanced MPE, CD8⁺ T cells displayed reduced IFN-γ, TNF-α, Granzyme B, Perforin, FasL, and Ki-67, but upregulated PD-1 and CTLA-4 relative to early stage. Similarly, Th1 cells showed decreased IFN-γ, TNF-α, and IL-2 production along with reduced Ki-67 expression. Advanced-stage M2 macrophages exhibited lower MHC-II levels and impaired phagocytosis, but higher PD-L1 and IL-10 production, while neutrophils showed reduced TNF-α release and phagocytic activity. Conclusions Our findings characterize the temporal immune dynamics associated with MPE progression in a mouse model, revealing a transition from an early immunostimulatory state to a late immunosuppressive state. This study enhances our understanding of MPE immunopathogenesis and provides a foundation for developing precise, stagespecific therapeutic strategies.

AIM To study the chemical constituents from Pyrolae Herba and their activity against pathogenic bacteria associated with sports injuries.METHODS Silica gel and Sephadex LH-20 were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antibacterial activity was detected by filter paper method.RESULTS Twenty five compounds were isolated and identified as manuleoside H(1),5-hydroxy-3,7-dimethoxy-4,-methyl flavone(2),5-hydroxy-3,7,4'-methoxy-flavone(3),5-hydroxy-7,3',4'-trimethoxy-flavone(4),5,7-dihydroxy-3-methoxy-flavone(5),7.5,4'-trihydroxyflavone(6),ethyl vanillate(7),clovandiol(8),hydroxydihydrobovolide(9),zanthopyranone(10),eugenin(11),berchemol(12),3,7-dimethoxy-5,3',4'-trihydroxyflavone(13),2,5-dimethyl-7-hydroxychromone(14),rutalinium(15),formononetin(16),8-hydroxy-4',7-dimethoxy-isoflavones(17),isoline(18),dearabinosyl pneumonanthoside(19),3,5-dihydroxycinnamic acid(20),2,4,6-trihydroxyacetophenone(21),p-isopropyl benzoic acid(22),geraldone(23),ethylsyringin(24),4-hydroxyphenylethyl-2'-hydroxypropionate(25).Compounds 8,10,12,16 and 24 showed bacteriostatic zone on pathogenic bacteria susceptible to sports injuries.CONCLUSION All the compounds are isolated from Pyrolae Herba for the first time.Compounds 8,10,12,16 and 24 have inhibitory activity against pathogenic bacteria susceptible to sports injuries.

Aim To explore the therapeutic effects of resveratrol(Res)on hepatic inflammation and oxida-tive stress in rheumatoid arthritis(RA),and to eluci-date the relationship of the regulatory mechanism of the Nrf2/Keap1 signaling pathway in it.Methods A mouse model of arthritis was induced using chicken type Ⅱ collagen in combination with complete Freund's adjuvant,and Res was administered by tube feeding for treatment.Serum liver function indices and levels of hepatic inflammation and oxidative stress were detected in mice.An in vitro cellular model of hepatic inflam-mation and oxidative stress was established by treating mouse primary hepatocytes(MPHs)with TNF-α(5μg·L-1),cell proliferation inhibition was detected by CCK-8,and inflammation and oxidative stress-relat-ed indices were detected by protein blotting.The in-trinsic mechanisms by which Res attenuated hepatic in-flammation and oxidative stress in rheumatoid arthritis were explored by treating MPHs with Nrf2 inhibitor and Keap1 overexpression plasmid.Results Res signifi-cantly reduced the levels of inflammation and oxidative stress in hepatic tissues of collagen-induced arthritis mice as well as TNF-α-treated MPHs,and activated the Nrf2/Keap1 signaling pathway.Inflammation and oxidative stress levels in MPHs were exacerbated by the use of Nrf2 inhibitors and Keap1 overexpression,which promoted apoptosis.Conclusion Res attenuates he-patic inflammation and oxidative stress in rheumatoid arthritis via the Nrf2/Keap1 pathway.

Aim To explore the therapeutic effects of resveratrol(Res)on hepatic inflammation and oxida-tive stress in rheumatoid arthritis(RA),and to eluci-date the relationship of the regulatory mechanism of the Nrf2/Keap1 signaling pathway in it.Methods A mouse model of arthritis was induced using chicken type Ⅱ collagen in combination with complete Freund's adjuvant,and Res was administered by tube feeding for treatment.Serum liver function indices and levels of hepatic inflammation and oxidative stress were detected in mice.An in vitro cellular model of hepatic inflam-mation and oxidative stress was established by treating mouse primary hepatocytes(MPHs)with TNF-α(5μg·L-1),cell proliferation inhibition was detected by CCK-8,and inflammation and oxidative stress-relat-ed indices were detected by protein blotting.The in-trinsic mechanisms by which Res attenuated hepatic in-flammation and oxidative stress in rheumatoid arthritis were explored by treating MPHs with Nrf2 inhibitor and Keap1 overexpression plasmid.Results Res signifi-cantly reduced the levels of inflammation and oxidative stress in hepatic tissues of collagen-induced arthritis mice as well as TNF-α-treated MPHs,and activated the Nrf2/Keap1 signaling pathway.Inflammation and oxidative stress levels in MPHs were exacerbated by the use of Nrf2 inhibitors and Keap1 overexpression,which promoted apoptosis.Conclusion Res attenuates he-patic inflammation and oxidative stress in rheumatoid arthritis via the Nrf2/Keap1 pathway.

The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

AIM To study the chemical constituents from Pyrolae Herba and their activity against pathogenic bacteria associated with sports injuries.METHODS Silica gel and Sephadex LH-20 were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antibacterial activity was detected by filter paper method.RESULTS Twenty five compounds were isolated and identified as manuleoside H(1),5-hydroxy-3,7-dimethoxy-4,-methyl flavone(2),5-hydroxy-3,7,4'-methoxy-flavone(3),5-hydroxy-7,3',4'-trimethoxy-flavone(4),5,7-dihydroxy-3-methoxy-flavone(5),7.5,4'-trihydroxyflavone(6),ethyl vanillate(7),clovandiol(8),hydroxydihydrobovolide(9),zanthopyranone(10),eugenin(11),berchemol(12),3,7-dimethoxy-5,3',4'-trihydroxyflavone(13),2,5-dimethyl-7-hydroxychromone(14),rutalinium(15),formononetin(16),8-hydroxy-4',7-dimethoxy-isoflavones(17),isoline(18),dearabinosyl pneumonanthoside(19),3,5-dihydroxycinnamic acid(20),2,4,6-trihydroxyacetophenone(21),p-isopropyl benzoic acid(22),geraldone(23),ethylsyringin(24),4-hydroxyphenylethyl-2'-hydroxypropionate(25).Compounds 8,10,12,16 and 24 showed bacteriostatic zone on pathogenic bacteria susceptible to sports injuries.CONCLUSION All the compounds are isolated from Pyrolae Herba for the first time.Compounds 8,10,12,16 and 24 have inhibitory activity against pathogenic bacteria susceptible to sports injuries.

Objective To analyze the innovative model and effectiveness of the four-level prevention of stroke in Nantong Medical and Prevention Collaborative Center for Cardiovascular and Cerebrovascular Disease,so as to provide new practical experience for the prevention and treatment of cerebrovascular disease.Methods In practice,Nantong city has explored the"53343"mode of collaborative treatment and prevention of cardiovascular and cerebrovascular diseases,and innovatively integrated this mode into four-level prevention to form a practice mode of zero-level guidance,first-level monitoring,second-level coordination and three-level leading.Data from the China Health Statistics Yearbook from 2011 to 2022 and the monitoring data of cardiovascular and cerebrovascular events in Nantong City on the Jiangsu Provincial Chronic Disease Management Information Platform from 2017 to 2023 were collected and processed by using Excel and SPSS 24.0,to analyze the changes of stroke surveillance data and effectiveness in Nantong City after applying the model.Results According to the monitoring data of cardiovascular and cerebrovascular events in Nantong City from the Jiangsu Provincial Chronic Disease Management Information Platform,despite the continuous increase in the number of monitored individuals over five years,the monitoring data for stroke from the Nantong Medical and Prevention Collaborative Center for Cardiovascular and Cerebrovascular Disease witnessed a decline in the number of monitored individuals for the first time in 2022.The stroke monitoring data from the same monitoring agency showed a more stable fluctuation trend compared to the number of national stroke patients discharged from 2017 to 2021.We established a coherent health service operation mechanism,cultivated 14 interdisciplinary talents,held a special continuing education class on stroke,strengthened the in-depth collaboration between clinical and public health,and provided a practical and innovative model for effective prevention and treatment of stroke.Conclusions The four-level prevention based on the medical and preventive collaboration center for cardiovascular and cerebrovascular diseases can achieve process and cycle health management,effectively slow down the upward trend of stroke,and improve the ability and level of scientific research and teaching.

BACKGROUND:Estrogen receptor α can act as an upstream protein to regulate the expression and phosphorylation level of adenosine monophosphate-activated protein kinase(AMPK).Activation of the estrogen receptor α-AMPK signaling pathway promotes osteogenic proliferation and differentiation.OBJECTIVE:To explore the molecular mechanism of estrogen receptor α regulating AMPK and its effect on osteoblast proliferation and differentiation at osteoblast cell line and molecular biology levels.METHODS:(1)The passaged MC3T3-E1 mouse embryonic osteoblasts were divided into three groups:blank control group,mock group(transfected with pCDNA3.1 control plasmid),and estrogen receptor α group(transfected with pCDNA3.1-estrogen receptor α overexpression plasmid),and RT-qPCR and western blot methods were used to detect the hepatic kinase B1,CaMKKβ,and AMPKα1 mRNA,protein and phosphorylation levels.(2)ChIP-qPCR was used to demonstrate that estrogen receptor α interacts with the hepatic kinase B1 promoter.Dual luciferase assay was used to demonstrate that estrogen receptorα interacts with the hepatic kinase B1 promoter region to activate its transcriptional expression.(3)The cells were divided into three groups:mock+shNC group,estrogen receptor α+shNC group,and estrogen receptor α+shLKB1 group.Changes in the expression levels of hepatic kinase B1,phosphorylated hepatic kinase B1,and phosphorylated AMPKα1 proteins in the cells were detected by western blot.(4)The cells were divided into four groups:mock group,estrogen receptor α group,estrogen receptor α+5 μmol/L Compound C(AMPK inhibitor)group,and estrogen receptor α+10 μmol/L Compound C group.The expression of proteins related to the AMPK signaling pathway and related to osteogenesis and osteoinductivity were detected by western blot method.Cells were transfected for 24 hours and then subjected to osteogenic induction for 14 days.Alkaline phosphatase staining was performed and cell viability in each group was detected.Mineralized nodule formation was detected by alizarin red staining at 21 days of osteogenic induction.(5)The cells were transfected and pretreated with different concentrations of AMPK inhibitor in corresponding groups,and cell viability was detected by cell counting kit 8.RESULTS AND CONCLUSION:(1)Estrogen receptor α activates the AMPK signaling pathway in MC3T3-E1 cells.(2)Estrogen receptor α promotes liver kinase B1 transcription and mediates AMPK signaling pathway activation.(3)Estrogen receptor α promotes the proliferation and differentiation of MC3T3-E1 cells by activating the AMPK signaling pathway,and the expression of AMPKα1,p-AMPKα,osteoprotegerin,osteopontin,and Runx2 proteins was down-regulated under the intervention of AMPK inhibitor,and the viability of osteoblasts was decreased.(4)To conclude,estrogen receptor α activates the AMPK signaling pathway by acting on liver kinase B1 promoter,promotes osteoblast proliferation and osteogenic differentiation,and prevents osteoporosis.

OBJECTIVE To carry out regular monitoring of carbapenem-resistant Klebsiella pneumoniae(CRKP)contamination status in the environment of intensive care unit(ICU)and take targeted prevention and control measures so as to reduce the incidence of hospital-associated infections with multidrug-resistant organisms(MDROs).METHODS The surfaces of surroundings of the patients who were colonized and infected with CRKP in the ICU of grade A tertiary hospital of Shanghai and the hands of relevant staff were sampled by stages from Jan 1,2021 to Jun 30,2024.The distribution of the CRKP strains in the surroundings were analyzed according to the locations positive for CRKP,and the disinfection measures were accordingly and continuously modified.The trend of isolation rate of CRKP strains from the ICU patients was analyzed during the time period when the measures were implemented.RESULTS Totally 266 environmental samples were collected during the baseline period(from Jan.1 2021 to Dec.31 2021),265 during intervention period(from Jan.1 2022 to Dec.31 2023),274 during con-solidation period(from Jan.1 to Jun.30 2024);the isolation rates of the CRKP strains were 4.51％,4.91％and 3.65％,respectively.The isolation rate of the strains was highest from the bed unit(10.40％),followed by the article for public use(6.74％),articles used by health care workers(2.98％)and diagnosis and treatment arti-cles(1.91％).The isolation rate of CRKP of the patients was 24.75％during the baseline period,15.48％during the intervention period,5.69％during the consolidation period,showing a continuously downward trend(x2=30.330,P＜0.001).CONCLUSION It is necessary to regularly carry out the environmental monitoring of CRKP strains,seek for the weak links of environmental disinfection and implement the intensified prevention and control measures so as to reduce the incidence of CRKP infection,which may provide theoretical bases for effective control of the CRKP strains.

Objective To explore the risk factors and intervention countermeasures for prolonged hospital stay in children with bronchopneumonia.Methods A retrospective analysis was conducted on the clinical data of 152 children with bronchopneumonia admitted to Zhoushan Women and Children Hospital from April 2022 to June 2024.The children were divided into prolonged group(hospital stay ≥ 10 days,53 cases)and non-prolonged group(hospital stay＜10d,99 cases)based on whether their hospital stay was prolonged.Multivariate Logistic regression analysis was used to determine the independent risk factors for prolonged hospital stay in children with bronchopneumonia.Pearson correlation analysis was used to determine the correlation between independent risk factors and prolonged hospital stay of the children.Results Univariate analysis showed that age,disease severity,duration of fever after admission,duration from onset to hospitalization,mycoplasma antibody,sputum culture,pneumonia and other related vaccination,and basic diseases were all influencing factors for prolonged hospital stay in children with bronchopneumonia(P＜0.05).Multivariate Logistic regression analysis showed that age,disease severity,and duration of fever after admission were all independent influencing factors for prolonged hospital stay in children with bronchopneumonia(P＜0.05).Age was negatively correlated with the prolongation of hospital stay(r=-0.285,P＜0.001).The disease severity and duration of fever after admission were positively correlated with the prolongation of hospital stay(r=0.312,0.458,P＜0.001).Conclusion Young age,severe condition and long duration of fever after admission are all independent risk factors for prolonged hospital stay in children with bronchopneumonia.