Objective To analyze the expression and detection significance of community acquired respiratory distress syndrome(CARDS)toxin in bronchoalveolar lavage fluid(BALF)in children with severe Mycoplasma pneumoniae pneumonia(SMPP).Methods The BALF samples of children diagnosed with Mycoplasma pneumoniae pneumonia(MPP)were collected from the Department of Pulmonology,Children's Hospital Affiliated to Capital Institute of Pediatrics from August 2021 to April 2022,the relative expression of CARDS toxins was determined by quantitative realtime polymerase chain reaction(qPCR).The patients were divided into SMPP group and non-SMPP group(control group).The relative expression of CARDS toxin and other clinical indicators were compared between the two groups.The risk factors of SMPP were analyzed.The predictive value of the indicators was analyzed using receiver operating characteristic(ROC)curves.Results ①The relative expression level of CARDS toxin(Z=-6.151),D-dimer(Z=-5.102)and lactate dehydrogenase(LDH)(Z=-6.337)in SMPP group were significantly higher than those in control group(P<0.001).The white blood cell(Z=-2.155),neutrophil ratio(t=2.988)and C-reactive protein(CRP)(Z=-3.360)were higher than those in control group(P<0.05).②The relative expression of CARDS toxin was positively correlated with serum LDH(r=0.258,P<0.05).③Binary Logistic regression analysis showed that fever time(OR=1.271,95% CI:1.079-1.497),LDH(OR=1.011,95% CI:1.005-1.017)and CARDS toxin(OR=6.210,95% CI:2.646-14.577)were risk factors for SMPP.Conclusion CARDS toxin was closely related to MPP disease,and was significantly increased in SMPP group.CARDS toxin was one of the independent risk factors for SMPP and positively correlated with serum LDH level.It can be used as a good indicator to evaluate the condition,combined with fever time and LDH has clinical significance.

Objective To establish a method for inducing mouse bone marrow monocytes differentiating into Kupffer cells in vitro,for the study of Kupffer cells.Methods Monocytes were separated from mouse bone marrow and induced by transforming growth factor 1(TGF-β1),macrophage colony-stimulating factor(M-CSF)and delta-like protein 4(DLL4).Induced monocyte-derived Kupffer cells(iMoKCs)were identified by real-time fluorescent quantitative polymerase chain reaction(qPCR),immunofluorescence staining and flow cytometry.Immunofluorescence staining and flow cytometry were used to evaluate the phagocytic capacity of iMoKCs.Cell counting kit-8(CCK-8)was used to evaluate the proliferation capacity.Results Bone marrow monocytes were isolated,after 24 h combined induction,iMoKCs express C-type lectin domain family 4,member f(CLEC4F),C-type lectin domain family 1,member b(CLEC1B)and V-set and immunoglobulin domain containing 4(VSIG4)at mRNA level,and protein CLEC4F.Phagocytic function of iMoKCs were detected by flow cytometry,and nearly 80% of total iMoKCs show bioparticle acceptance.The number of iMoKCs were detected by CCK-8,the counts were 1.4,2.0 and 2.9 times of the initial number(0 h)at 24,48 and 72 h.Conclusion These cells display the immunophenotype of Kupffer cells and show the ability of phagocytosis and proliferation induction through M-CSF,TGF-β1 and DLL4,which could replace primary Kupffer cells for in vitro research.

Objective To evaluate the transition rules of normal body mass index(BMI),overweight and metabolic syndrome(MetS)in patients with major depressive disorder(MDD).Methods Patients with MDD who had multiple admission records between Jan 2016 and Nov 2021 in Beijing Anding Hospital,Capital Medical University were included.Based on the overweight and metabolic syndrome status assessed at each admission,the patients were categorized into three states:normal BMI,overweight and metabolic syndrome.A multi-state Markov model was used to analyze the transition intensity and transition frequency between three states and the influence of covariates on transitions.Results A total of 892 records of 398 subjects were included,with a median age of 56 years old and 31.4% males.The median follow-up period was 40 months.The multi-state model showed that there were 494 transitions between the three states,of which 5.1% moved from normal BMI to overweight and 5.5% moved from overweight to MetS.The intensity of transition was the highest from overweight to MetS,9.52 times greater than overweight to normal BMI.After 48.53 months,MDD patients with normal BMI began to transition to MetS.For overweight MDD patients,the transition to MetS started after 8.77 months.MDD patients with normal BMI or overweight had 31.4% and 50.4% probabilities of developing Mets after 36 months.For MDD patients comorbid with MetS,the probability of staying at MetS was 51.2% after 36 months.Multivariate analysis showed that being unmarried was a risk factor against developing overweight in normal BMI MDD patients,while a higher level of education was a protective factor against developing MetS in overweight MDD patients.Conclusion MDD patients exhibited a higher intensity and risk of developing MetS,and it is not easy to reverse MetS,suggesting that BMI management and MetS intervention should be strengthened in MDD patients.

Objective To evaluate clinical effect of remote ischemic conditioning(RIC)in patients with spontaneous intracerebral hemorrhage(sICH).Methods Sixty-four patients with sICH who were diagnosed and admitted to the Department of Neurosurgery,the Sixth Medical Center of PLA General Hospital from January 2023 to May 2024 were selected as research subjects.Following the principle of matching baseline characteristics between groups,a computerized random grouping program was used to divide them into an observation group of 32 cases and a control group of 32 cases.Patients in the control group received standard basic medication according to the'Chinese Clinical Management Guidelines for Cerebrovascular Diseases',while the observation group received RIC treatment in addition to the control group's treatment,with a course of 14 d.The changes in National Institutes of Health Stroke Scale(NIHSS)scores and Barthel index(BI,daily living ability score table)on the day of admission and after 14 d of treatment,the changes in hematoma volume on computed tomography(CT)imaging,and the incidence of adverse events were compared between the two groups.Results At admission,the difference is not statistically significant in NIHSS scores,BI,and CT imaging hematoma volume between the two groups(P>0.05).After 14 d of treatment,the NIHSS scores of both groups were reduced compared to before treatment,and the scores of the observation group were lower than those of the control group(P<0.05);the BI of both groups was increased compared to before,and the scores of the observation group were higher than those of the control group(P<0.05);there were no significant differences in CT imaging hematoma volume and the incidence of adverse events between the two groups(P>0.05).Conclusion Continuous RIC treatment for 14 d is safe and well-tolerated in patients with spontaneous intracerebral hemorrhage and can effectively improve the neurological function of patients.

Objective To introduce the concepts and principles of the win ratio method and to analyze it in the context of a case study of a clinical trial in cerebrovascular disease.Methods Based on the study of clopidogrel with aspirin in high risk patients with acute non disabling cerebrovascular events 2,and key prognostic factors,the outcome events were defined sequentially as ① time to death within 90 d,② time to recurrence of ischemic stroke within 90 d,③ time to moderate-to-severe hemorrhage within 90 d.Using clopidogrel combined with aspirin as the reference group,the winning ratio(Rw)of ticagrelor combined with aspirin was analyzed by the win ratio method,and the 95% confidence interval(CI)of Rw was estimated by the Bootstrap method and compared with the hazard ratio(HR)calculated by the competing risk model.Results When only fatal events were considered,the win ratio method suggested that the ticagrelor group was significantly better than the clopidogrel group,Rw=2.00(95% CI:1.52-2.47),and after stepwise inclusion of ischemic stroke and moderate-to-severe hemorrhage recurrence,the win ratio method yielded a value of 1.29(95% CI:1.25-1.57),and the HR value from Fine and Gray competing risk regression was 0.78(95% CI:0.65-0.95),both of which indicated that the efficacy of the ticagrelor group was superior to that of the clopidogrel group.Conclusion The win ratio method can be used to analyze clinical trials with composite endpoints after prioritizing multiple outcome variables,showing the advantages of win ratio and its promising application in cerebrovascular disease research.

Acute ischemic stroke(AIS)is associated with high mortality and disability rates,presenting a substantial challenge to global public health challenge.Intravenous thrombolysis(IVT)is recognized as a cornerstone of early AIS treatment and is recommended as the standard therapeutic approach by both national and international guidelines.However,the clinical efficacy of IVT remains suboptimal due to several limitations,including a narrow therapeutic time window and the inevitable activation of the coagulation system and platelet aggregagation during thrombolysis.These factors may contribute to adverse outcomes such as early neurological deterioration(END)and vascular re-occlusion.Antiplatelet therapy(APT),which inhibits platelet aggregations,reduces microthrombus formation,and stabilizes the vascular endothelium with multifaceted mechanisms,has emerged as a promising adjunctive strategy to IVT,offering potential synergistic effects.This review summarized the latest evidence from both domestic and international studies,focusing on the mechanisms of APT,recent clinical advancements in IVT combined with APT,and the safety and efficacy of APT administration at different time windows relative to IVT.Emphasis is placed on the influence of various antiplatelet agents,dosing regimens,and initiation timing on therapeutic outcomes,alongside a comprehensive evaluation in the context of current guideline recommendations and clinical practice.Current guidelines recommend initiating APT 24 h after IVT,following imaging confirmation to exclude the risk of intracranial hemorrhage.However,the efficacy and safety of earlier APT initiation remain inconclusive.Individualized treatment strategies,such as early administration of low-dose,short-acting APT or combination therapy in specific patient subgroups,may effectively balance therapeutic benefits and risks.The adjunctive use of APT in IVT holds promise for enhancing efficacy and improving clinical outcomes,but precise stratification of safety and efficacy is essential.Future research should focus on optimizing combination IVT and APT strategies through individualized patient profiling,appropriate drug selection,and dynamic imaging monitoring to achieve precision management in AIS treatment.

Objective To elucidate the functional role and underlying mechanisms of the ribosome biogenesis factor BMS1 in neuroblastoma(NB)cellular proliferation.Methods We utilized the R2 genomics analysis and visualization platform to analyze the correlation between BMS1 expression levels and clinical characteristics of NB children.The BMS1 mRNA level in three human neuroblastoma cells SK-N-BE(2),BE(2)-C,IMR-32 and two normal cells hTERT RPE-1,IMR-90 was detected by real-time quantitative polymerase chain reaction(RT-qPCR).Two distinct small interfering RNA(siRNA)sequences were used to target BMS1 mRNA in NB cells SK-N-BE(2)and BE(2)-C,with normal cells hTERT RPE-1 serving as controls.We used RT-qPCR to quantify the mRNA levels of BMS1 and two key neuroblastoma-associated molecules(MYCN and p53).After transfection with siRNA,cellular proliferation was detected by various experimental approaches:crystal violet staining,real-time cell analysis(RTCA),colony-forming unit assay and immunofluorescence.Results By analyzing two independent neuroblastoma clinical cohorts(GSE85047/NRC-283 and Westermann-144 datasets),it was found that the BMS1 mRNA level in MYCN-amplified NB was significantly higher than that in MYCN-non-amplified NB(P<0.05).Furthermore,the overall survival rate of NB children in the BMS1 high-expression group was decreased(P<0.05).Consistent with these clinical observations,the BMS1 mRNA level in NB cells SK-N-BE(2),BE(2)-C and IMR-32 was significantly higher than that in normal cells hTERT RPE-1,IMR-90(P<0.05).The targeted transient knockdown of BMS1 in NB cell lines SK-N-BE(2)and BE(2)-C resulted in decreased intracellular MYCN mRNA expression levels(P<0.05),significantly reduced cell proliferation capacity and colony-forming ability(P<0.05).Immunofluorescence revealed that the expression of Ki-67,a proliferation marker,was decreased(P<0.05).At the molecular level,RT-qPCR showed that the p53 mRNA level was significantly elevated in the BMS1-knockdown groups(si BMS1-1#and si BMS1-2#)compared with the control group(P<0.05).However,transient knockdown of BMS1 had no significant impact on the proliferative capacity of normal cells hTERT RPE-1.Conclusion BMS1 expression was up-regulated in MYCN-amplified NB and negatively correlated with the prognosis of the NB children.Mechanistically,interfering with BMS1 expression may transcriptionally activate p53 in NB cells,thereby inhibiting their proliferative ability,while having minimal impact on normal cells growth kinetics.These findings suggest that BMS1 serves as an important proliferation driver in NB and is expected to be a promising therapeutic target for NB children,particularly MYCN-amplified pediatric patients.

The integration of artificial intelligence(AI)into medical practice has significantly impacted the field of cerebrovascular disease.AI algorithms are increasingly being employed to enhance the diagnosis and management of cerebrovascular conditions.However,the clinical application and accuracy of these AI tools require further rigorous evaluation.This review probes into the current applications of AI in diagnosis and decision-making in cerebrovascular disease,and explores the potential and challenges associated with their implementation.

Objective To clarify the effect of unilateral activation of the dopamine type I receptor medium-sized multi-spiny neurons(D1-MSN)in the dorsal striatum of mice on speed.Methods The transgenic animals were combined with optogenetic experiments to specifically activate the D1-MSN in the dorsal striatum of mice at different frequencies and to analyze the rotational behavior and speed of mice when stimulating the D1-MSN.Results Unilateral activation of D1-MSN induces contralateral rotational behavior in mice and either increases or decreases speed.The mechanisms by which different frequencies affect the speed of mice differently.As the frequency of stimulus increased,the contralateral rotational behavior of the mice increased.Unilateral stimulus of D1-MSN increased speed and induced contralateral rotational behavior,and the rotational behavior increased with increasing stimulus frequency.In the experiment where D1-MSN stimulus did not induce rotational behavior,it was found that 5 Hz stimulus still induced an increase in speed,but 15 Hz and 25 Hz stimulus did not induce an increase in speed.Further analysis of the pre-stimulus locomotor state of the mice showed that 5 Hz,15 Hz and 25 Hz stimulus increased speed when the average speed before stimulus was less than 5 cm/s.However,15 Hz stimulus decreased the speed when the average speed before stimulus was greater than 5 cm/s.Conclusion Unilateral activation of D1-MSN in the dorsal striatum modulates speed and induces contralateral rotational behavior in mice,and is related to the frequency of stimulus and the locomotor state of the mice before stimulus.

Objective To develop a Nomogram clinical prediction model for the pathological occurrence of extraprostatic extension(EPE)after radical prostatectomy in prostate cancer patients,using simplified site-specific magnetic resonance imaging(MRI)indicators and other clinical parameters.Methods A total of 181 prostate cancer patients[mean age(69.0±7.3)years]who underwent radical prostatectomy were included.These patients had received 3-Tesla multi-parametric magnetic resonance imaging(3-T mpMRI)within 6 months prior to surgery.Based on mpMRI measurements[capsular contact length(CCL)>15 mm,capsular bulging/irregularities,diameter of index lesion(dIL),and evident extraprostatic extension(eEPE)],the dIL?sEPE grading system was derived.The optimal cut-off value of dIL(denoted as dIL)was determined using the Youden J index,and categorized it into a binary variable.A Logistic regression model was established based on the dIL?sEPE grading and clinical scores.The predictive performance of clinical indicators,MRI indicators,and combined clinical and MRI indicators were compared.Finally,a clinical prediction model integrating both clinical and MRI data was developed.Results Pathological EPE was found in 46 out of 181 cases(25.4% ).A Nomogram prediction model for EPE was established with a combination of the dIL?sEPE grading and clinical indicators.Conclusion The combination of dIL?sEPE grading with clinical indicators accurately predicts extracapsular extension in prostate cancer.The Nomogram model that established,based on MRI imaging characteristics and clinical indicators has good performance and is easy to use.It is beneficial to stratifying management for prostate cancer patients,and it provides valuable guidance for patients suitable for nerve-sparing surgery.