Objective: To investigate whether membrane vesicles (MVs) of Streptococcus mutans （S.mutans） contain autoinducer-2 (AI-2) and to preliminarily explore the effects of these MVs on the growth and biofilm formation of S. mutans. Methods: MVs were isolated from the S. mutans UA159 strain using differential centrifugation. The isolated MVs were characterized by nanoparticle tracking analysis for particle size and concentration and observed by transmission electron microscopy. The presence of AI-2 was identified using the Vibrio harveyi BB170 bioluminescence assay: the BB170 diluent was supplemented with AB medium (control group), MV extract (MVs group), pre-ultrafiltration supernatant (Sup group), or post-ultrafiltration supernatant (Sup-af group). The effects of MVs on growth and biofilm formation were assessed using the S.mutans UA159 strain or a luxS deletion mutant as the control group, compared with experimental groups stimulated with gradient concentrations of MVs (MVs-2.0E+7, MVs-2.0E+8, and MVs-2.0E+9 groups). Growth curves, MTT assay, and colony-forming unit (CFU) counts were used to determine changes in growth capacity. Biofilm formation was evaluated using crystal violet staining, confocal laser scanning microscopy, and the anthrone method for polysaccharide quantification. Results: Enriched S. mutans MVs were successfully obtained, with an average particle size of approximately 94.19 nm and a concentration of 1.87E+11 particles/mL. The bioluminescence assay showed that the luminescence intensity of the Sup group was higher than that of the Sup-af group, and the MVs group exhibited higher intensity than the control group. Assessments via growth curves, MTT assay, and CFU counts indicated no significant differences in the growth capacity of the various S. mutans strains after treatment with different concentrations of MVs. Crystal violet staining quantification and confocal laser scanning microscopy observations revealed that high-concentration MV treatment (2.0E+9 particles/mL group) resulted in lower biofilm mass compared to the control. The anthrone method showed that the production of both water-soluble and water-insoluble polysaccharides was significantly lower in the high-concentration MV group than in the control. Conclusion S. mutans MVs contain the quorum sensing signal molecule AI-2. These MVs do not significantly affect the growth of S. mutans, but they can regulate biofilm formation and exhibit an inhibitory effect at high concentrations.

Polycystic ovary syndrome （PCOS）， a common reproductive endocrine disorder in women， is one of the leading causes of ovulatory infertility in women of reproductive age. Due to its heterogeneous etiology， complex symptoms， and challenging treatment， PCOS has become a focal point of research in gynecological and reproductive medicine globally. The pathogenesis of PCOS is complex and may involve regulatory mechanisms such as inflammatory responses， oxidative stress， and cellular autophagy. Crown-like structures （CLSs） refer to pro-inflammatory microenvironments formed by macrophages engulfing adipocytes. The inflammatory disorders induced by CLSs are one of the key factors contributing to the development of PCOS and its complications. Current studies have indicated that the obese status in PCOS accelerates the formation of CLSs， and the density of CLSs can predict the progression of metabolic disorders and influence the outcomes of various metabolic diseases. Traditional Chinese Medicine （TCM） offers the unique advantages of a holistic view， four diagnostic methods， and syndrome differentiation and treatment to ameliorate the symptoms and signs of PCOS through multiple levels， pathways， and targets. Although studies on the mechanisms of metabolic diseases and CLS formation have been reported in China and abroad， there is still a lack of literature on the correlation between CLSs and PCOS， as well as reviews on TCM interventions targeting CLSs for treating this disease. Therefore， this paper summarized the correlation between obese PCOS and CLSs and reviewed recent studies on TCM interventions based on CLS formation （adipose tissue-macrophage inflammatory crosstalk） in the treatment of obese PCOS， aiming to provide new research perspectives for the prevention and treatment of PCOS using TCM.

Polycystic ovary syndrome （PCOS）， a common reproductive endocrine disorder in women， is one of the leading causes of ovulatory infertility in women of reproductive age. Due to its heterogeneous etiology， complex symptoms， and challenging treatment， PCOS has become a focal point of research in gynecological and reproductive medicine globally. The pathogenesis of PCOS is complex and may involve regulatory mechanisms such as inflammatory responses， oxidative stress， and cellular autophagy. Crown-like structures （CLSs） refer to pro-inflammatory microenvironments formed by macrophages engulfing adipocytes. The inflammatory disorders induced by CLSs are one of the key factors contributing to the development of PCOS and its complications. Current studies have indicated that the obese status in PCOS accelerates the formation of CLSs， and the density of CLSs can predict the progression of metabolic disorders and influence the outcomes of various metabolic diseases. Traditional Chinese Medicine （TCM） offers the unique advantages of a holistic view， four diagnostic methods， and syndrome differentiation and treatment to ameliorate the symptoms and signs of PCOS through multiple levels， pathways， and targets. Although studies on the mechanisms of metabolic diseases and CLS formation have been reported in China and abroad， there is still a lack of literature on the correlation between CLSs and PCOS， as well as reviews on TCM interventions targeting CLSs for treating this disease. Therefore， this paper summarized the correlation between obese PCOS and CLSs and reviewed recent studies on TCM interventions based on CLS formation （adipose tissue-macrophage inflammatory crosstalk） in the treatment of obese PCOS， aiming to provide new research perspectives for the prevention and treatment of PCOS using TCM.

Objective To analyze the monitoring of healthcare-associated infection(HAI)in the intensive care unit(ICU)over the past 12 years based on Joinpoint regression model,and evaluate the trend changes and relevant fac-tors of HAI incidence.Methods ICU patients in a tertiary first-class hospital from January 2012 to December 2023 were selected and performed prospective monitoring.Trend changes of HAI incidence and the correlation with con-sumption of hand hygiene products as well as HAI management measures were analyzed.Results From 2012 to 2023,6 929 ICU patients were included in the monitoring,543 patients had 655 episodes of HAI,with incidence and case incidence of HAI being 7.84％ and 9.45％,respectively.The average severity of the disease was 3.62,and the adjusted HAI incidence was 2.17％.The daily incidence of ventilator-associated pneumonia(VAP),cathe-ter-associated urinary tract infection(CAUTI),and central line-associated bloodstream infection(CLABSI)were 6.19‰,3.45‰,and 1.23‰,respectively.The consumption of hand hygiene products was 122.98 mL/bed-day.The compliance rate and correct rate of hand hygiene were 90.63％and 90.46％,respectively.From 2012 to 2023,incidence of HAI(51.29％vs 4.39％),case incidence of HAI(72.41％vs 4.94％),the adjusted incidence of HAI(15.98％vs 1.04％),daily incidence of VAP(22.50‰ vs 4.33‰),daily incidence of CAUTI(14.23‰ vs 1.64‰),and daily incidence of CLABSI(10.60‰ vs 0.20‰)all decreased significantly(all P＜0.05).Both con-sumption of hand hygiene products(75.16 mL/bed-day vs 147.35 mL/bed-day)and correct rate of hand hygiene(85.00％vs 90.28％)increased significantly(both P＜0.05).A total of 1 946 pathogens were detected,with an increase in the proportion of Staphylococcus aureus(1.30％ vs 9.57％)and a decrease in the proportion of fungi(11.04％vs 1.74％).The daily consumption of hand hygiene products negatively correlated with the incidence of HAI,the case incidence of HAI,as well as the daily incidence of CAUTI and CLABSI(all P＜0.05).Incorpora-ting HAI real-time monitoring system and HAI management into performance assessment could decrease HAI-rela-ted incidence(P＜0.05).Conclusion HAI-related incidence presents a downward trend.Scientific and comprehen-sive HAI prevention and control management measures such as healthcare workers'hand hygiene management,on-line HAI real-time monitoring system,and incorporating HAI management into performance assessment can de-crease HAI-related incidence and promote the improvement of medical quality.

Objective To analyze the monitoring of healthcare-associated infection(HAI)in the intensive care unit(ICU)over the past 12 years based on Joinpoint regression model,and evaluate the trend changes and relevant fac-tors of HAI incidence.Methods ICU patients in a tertiary first-class hospital from January 2012 to December 2023 were selected and performed prospective monitoring.Trend changes of HAI incidence and the correlation with con-sumption of hand hygiene products as well as HAI management measures were analyzed.Results From 2012 to 2023,6 929 ICU patients were included in the monitoring,543 patients had 655 episodes of HAI,with incidence and case incidence of HAI being 7.84％ and 9.45％,respectively.The average severity of the disease was 3.62,and the adjusted HAI incidence was 2.17％.The daily incidence of ventilator-associated pneumonia(VAP),cathe-ter-associated urinary tract infection(CAUTI),and central line-associated bloodstream infection(CLABSI)were 6.19‰,3.45‰,and 1.23‰,respectively.The consumption of hand hygiene products was 122.98 mL/bed-day.The compliance rate and correct rate of hand hygiene were 90.63％and 90.46％,respectively.From 2012 to 2023,incidence of HAI(51.29％vs 4.39％),case incidence of HAI(72.41％vs 4.94％),the adjusted incidence of HAI(15.98％vs 1.04％),daily incidence of VAP(22.50‰ vs 4.33‰),daily incidence of CAUTI(14.23‰ vs 1.64‰),and daily incidence of CLABSI(10.60‰ vs 0.20‰)all decreased significantly(all P＜0.05).Both con-sumption of hand hygiene products(75.16 mL/bed-day vs 147.35 mL/bed-day)and correct rate of hand hygiene(85.00％vs 90.28％)increased significantly(both P＜0.05).A total of 1 946 pathogens were detected,with an increase in the proportion of Staphylococcus aureus(1.30％ vs 9.57％)and a decrease in the proportion of fungi(11.04％vs 1.74％).The daily consumption of hand hygiene products negatively correlated with the incidence of HAI,the case incidence of HAI,as well as the daily incidence of CAUTI and CLABSI(all P＜0.05).Incorpora-ting HAI real-time monitoring system and HAI management into performance assessment could decrease HAI-rela-ted incidence(P＜0.05).Conclusion HAI-related incidence presents a downward trend.Scientific and comprehen-sive HAI prevention and control management measures such as healthcare workers'hand hygiene management,on-line HAI real-time monitoring system,and incorporating HAI management into performance assessment can de-crease HAI-related incidence and promote the improvement of medical quality.

Objective To investigate the impacts of sevoflurane combined with lung protective ventilation strategy on pulmonary ventilation function and lung compliance in obese patients undergoing laparoscopic weight loss surgery.Methods 60 obese patients underwent laparoscopic weight loss surgery were randomly divided into two groups.The control group was given lung protective ventilation intervention alone during anesthesia,and the study group was given sevoflurane inhalation anesthesia combined with lung protective ventilation intervention.Arterial blood was collected before tracheal intubation(T0),5 min after tracheal intubation(T1),40 min after tracheal intubation(T2)and 5 min after tracheal extubation(T3)for blood gas analysis.The pulmonary ventilation function and lung compliance of patients in the two groups were compared.Results Peak airway pressure(Ppeak)and plateau airway pressure(Pplat)at T2 were lower in the study group than those in the control group,and the differences were statistically significant(P<0.05);At T2 and T3 time points,the dynamic lung compliance(Cldyn)of the study group was higher than that of the control group,and the differences were statistically significant(P<0.05);7 days after surgery,the forced vital capacity(FVC)and forced expiratory volume in one second(FEV1)in the study group were higher than those in the control group,and the differences were statistically significant(P<0.05);At time points T1,T2 and T3,the levels of serum transforming growth factor-β1(TGF-β1),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the study group were lower than those in the control group(P<0.05);After surgery,the awakening time,spontaneous breathing recovery time,and extubation time in the study group were shorter than those in the control group,the number of adverse events during the recovery period was less than that in the control group,after awakening,the Ramsay score was lower than that in the control group(P<0.05).Conclusion The combination of sevoflurane and lung protective ventilation strategy can reduce inflammatory response,improve pulmonary ventilation function,and improve lung compliance in obese patients undergoing laparoscopic weight loss surgery,with good safety and fast postoperative recovery.

Objective This study aims to establish a non-puerperal mastitis (NPM) rat model by simulating hyperprolactinemia and immune-inflammatory states, and to evaluate its local inflammatory characteristics in the mammary gland, thereby laying the foundation for research on the diagnosis and treatment of this clinically challenging disease. Methods Twelve SPF-grade Wistar female rats were evenly divided into a control group and a model group. During the experiment, the control group received no experimental treatment or medication. The model group received daily subcutaneous injections of 100 mg/kg metoclopramide hydrochloride for 7 consecutive days. Serum prolactin (PRL) levels were measured using ELISA on the 10th, 20th, and 30th days after the first injection. After 7 days of injections, 200 μL of lactating SD rat milk was mixed with 200 μL of complete Freund's adjuvant to prepare an oil-in-water emulsion, which was administered by multiple subcutaneous injections into the back of the Wistar rats for the initial immunization. Seven days after the initial immunization, the emulsion was injected subcutaneously into the third, fourth, and fifth mammary glands for the final immunization. After the final immunization, the rats were observed for 28 days for changes in mammary gland appearance, and the size of mammary nodules was calculated. On the 3rd, 7th, 14th, and 28th days, hematoxylin-eosin (HE) staining was used to analyze mammary tissue morphology. Immunohistochemistry was employed to detect CD138 expression levels. ELISA was used to measure the levels of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) in mammary tissue to comprehensively assess the model. Results Rats in the model group exhibited mammary skin ulceration and foul odor at the ulcer sites. Palpation and ultrasound revealed the formation of mammary nodules. HE staining showed that on the 3rd day after the final immunization, normal ductal and lobular structures in the mammary glands disappeared, with significant infiltration of plasma cells. On the 7th day, ductal dilation, epithelial necrosis and detachment, and pronounced periductal plasma cell and lymphocyte (predominantly T-lymphocytes) infiltration were observed. On the 14th day, there was a proliferation of fibrofatty tissue, small blood vessels, and granulation tissue, with scattered plasma cells in the interstitium. By the 28th day, inflammatory cell infiltration and fibrous tissue proliferation were reduced, with granuloma formation. Serum PRL levels in the model group were significantly increased on the 10th day (P<0.05) and the 20th day (P<0.001). IL-6 and TNF-α levels in mammary tissue were higher in the model group compared to the control group on the 3rd, 7th, 14th, and 28th days (P<0.05). IL-1β levels were higher on the 3rd, 7th, and 14th days compared with the control group (P<0.01) but lower than the control group on the 28th day (P>0.05). iNOS levels were significantly higher on the 7th day after the final immunization (P<0.001). Conclusion A successful NPM model was established in rats, which exhibited typical pathological features such as local mammary masses, abscesses, ulcers, ductal dilation and plasma cell infiltration. This model can serve as a foundation for further research into the diagnosis and treatment of this clinically challenging disease.

Background::Pulmonary embolism (PE) is a severe and acute cardiovascular syndrome with high mortality among patients with autoimmune inflammatory rheumatic diseases (AIIRDs). Accurate prediction and timely intervention play a pivotal role in enhancing survival rates. However, there is a notable scarcity of practical early prediction and risk assessment systems of PE in patients with AIIRD.Methods::In the training cohort, 60 AIIRD with PE cases and 180 age-, gender-, and disease-matched AIIRD non-PE cases were identified from 7254 AIIRD cases in Tongji Hospital from 2014 to 2022. Univariable logistic regression (LR) and least absolute shrinkage and selection operator (LASSO) were used to select the clinical features for further training with machine learning (ML) methods, including random forest (RF), support vector machines (SVM), neural network (NN), logistic regression (LR), gradient boosted decision tree (GBDT), classification and regression trees (CART), and C5.0 models. The performances of these models were subsequently validated using a multicenter validation cohort.Results::In the training cohort, 24 and 13 clinical features were selected by univariable LR and LASSO strategies, respectively. The five ML models (RF, SVM, NN, LR, and GBDT) showed promising performances, with an area under the receiver operating characteristic (ROC) curve (AUC) of 0.962-1.000 in the training cohort and 0.969-0.999 in the validation cohort. CART and C5.0 models achieved AUCs of 0.850 and 0.932, respectively, in the training cohort. Using D-dimer as a pre-screening index, the refined C5.0 model achieved an AUC exceeding 0.948 in the training cohort and an AUC above 0.925 in the validation cohort. These results markedly outperformed the use of D-dimer levels alone.Conclusion::ML-based models are proven to be precise for predicting the onset of PE in patients with AIIRD exhibiting clinical suspicion of PE.Trial Registration::Chictr.org.cn: ChiCTR2200059599.

Objective To investigate and validate the expression profiles of Ppp3cb and Ppm1g through differential proteomic analysis of hippocampal tissue in NHE1 gene knockout mice with proteomic analysis.Method ① Six 2-week-old NHE1 knockout mice were selected as the model group,and 6 wild-type mice of the same age served as the control group,and their genotypes were detected by agar-gel electrophoresis.Open field test and forced swimming test were used to evaluate the behaviors of mice in the model group and control group,and epileptic seizure was graded according to Racine scoring.② Tandem mass spectrometry was employed to screen the differential proteins in the hippocampus tissues from the model group and the control group.Then the obtained differential proteins were annotated and enriched in the Gene Ontology(GO)database.Search tool for the retrieval of interesting genes(STRING)database was used to analyze protein-protein interaction(PPI)among different proteins.③ The transcriptional and translational levels of Ppp3cb and Ppm1g were detected by qPCR and Western blotting,respectively,and their expression levels in the tissues were observed with immunohistochemistry.Results ① NHE1 was not expressed in the model group.The mice of the model group had shorter total movement distance(P=0.007 3)and less crossing cells(P＜0.000 1)in open field test,and longer period of immobility in forced swimming test(P＜0.000 1)when compared with those from the control group.② When fold change ≥1.2 times and P＜0.05 were set as the significant threshold for differential expression,845 differentially expressed protein sites were detected in the hippocampus,among which 9 proteins(including Ppm1g)were up-regulated and 7 ones(including Ppp3cb)were down-regulated.Gene Ontology(GO)functional analysis showed that after NHE1 knockout,the most significant differences were observed in the concentration of molecular function(MF)related to protein serine/threonine phosphatase activity,concentration of cellular component(CC)related to the plasma membrane,and concentration of biological process(BP)related to negative regulation of biological processes and immune system processes.STRING analysis indicated that the differential proteins Ppp3cb and Slc9a1 directly acted,Ppm1g indirectly acted through Ppp3cb and Slc9a1,and Ppp3cb and Ppm1g interacted.③The transcriptional and translational levels of Ppp3cb were decreased,and its expression level was reduced in the tissues,while those of Ppm1g were increased,and its expression was elevated in the tissues(P＜0.05).Conclusion In the hippocampus of NHE1 gene knockout mice,the expression of differential protein Ppp3cb is down-regulated and that of Ppm1g is up-regulated,which provide a basis for further study on their involvement in the pathogenesis of epilepsy.

AIM To analyze the component composition of Xeriga-4 Powder,and to determine the contents of phellodendrine,chlorogenic acid,gardenoside,berberine,rutin and curcumin.METHODS The high performance liquid chromatography-Q-exactive orbitrap mass spectrometry(HPLC-Q-Exactive-MS)qualitative analysis was performed on a 35℃thermostatic Agilent ZORBAX SB-Aq column(4.6 mm×150 mm,5 μm),with the mobile phase comprising of methanol-0.1%formic acid flowing at 0.35 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning.High performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)quantitative analysis was performed on a 35℃thermostatic Shim-pack GIST-HP C18 column(2.1 mm×100 mm,3 μm),with the mobile phase comprising of methanol-0.1%formic acid flowing at 0.25 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning with multiple reaction monitoring mode.RESULTS Total 65 constituents were identified,containing 19 alkaloids,13 organic acids,13 flavonoids,7 curcumins,6 iridoids,4 fatty acids,2 aldehydes,and 1 amino acid.Six constituents showed good linear relationships within their own ranges(r≥0.999 1),whose average recoveries were 96.44%-102.37%with the RSDs of 2.05%-3.74%.CONCLUSION This study can provide a reference for the quality control for Xieriga-4 Powder.