To understand the epidemiological characteristics and antimicrobial resistance of Esche-richia coli in free-range Tibetan pig populations,fecal samples from these pigs were collected be-tween 2021 and 2023 for pathogen isolation and identification.The isolated strains underwent anal-ysis of pathogenicity categories,phylogenetic grouping,and antimicrobial susceptibility profiles,a-long with resistance gene detection and mouse pathogenicity testing.The results showed that 123 strains of E.coli were isolated from 132 fecal samples.Enteropathogenicity tests revealed that 42.28％were EAEC(52/123),7.32％were EPEC(9/123),and 3.25％were STEC/EHEC(4/123).Phylogenetic analysis indicated that 78.86％(97/123)belonged to group A,while 11.38％(14/123)belonged to group B1.Antimicrobial susceptibility tests for 22 drugs showed the highest resistance rate to amoxicillin at 98.37％(121/123),followed by sulfamethoxazole at 73.98％(91/123).Resistance rates for ampicillin,tetracycline,erythromycin,and trimethoprim ranged from 48.78％to 51.22％.Furthermore,78.86％(97/123)of the isolates were resistant to three or more antibiotics.The detection of 52 drug-resistance genes in 10 categories showed that 15 drug-resist-ance genes were detected,and the detection rate was 28.85％(15/52).Among individual resistance genes,tetA had the highest detection rate at 63.41％(78/123),followed by tetB and qnrS with de-tection rates of 48.78％(60/123)and 38.21％(47/123),respectively.This study demonstrates that EAEC is the predominant strain of E.coli in free-range Tibetan pigs,with groups A and B1 being the major phylogenetic groups.The isolates exhibited a high degree of multidrug-resistant pheno-types,and the detection rates of resistance genes were also high.E.coli carrying virulence genes cause different degrees of pathological changes in the mouse ileum.The research results have sig-nificant public health implications and provide theoretical basis for the prevention,treatment,and clinical medication of E.coli disease in Tibetan pigs.

Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.

OBJECTIVE To conduct the epidemiological survey for 4 patients with carbapenem-resistant Klebsiella pneumoniae(CRAB)that gathered to emerge in intensive care unit(ICU)within a short period so as to find out the sources of suspected CRAB.METHODS The clinical data and epidemiological history of the 4 patients with hospital-associated infection with CRAB who were treated in Shandong Provincial Hospital Affiliated to Shandong First Medical University from Mar.31,2023 to Apr.7,2023 were retrospectively analyzed.The whole genome second-generation sequencing was carried out for the CRAB strains isolated from the patients and their surround-ings,and the sources of the CRAB strains were traced by means of the whole genome sequencing.RESULTS The result of the epidemiological survey showed that the case 1,2,3 and 4 were close in spatial position and had the similar drug resistance spectrum,and there might be cross infection.With the combination of the result of genome analysis,it was found that the strain from the case 1 was brought from the community and was the evolutionary branch different from other CRAB strains causing nosocomial infection;the strains from the case 2,3 and 4 had the close original sources with the CRAB strains from surroundings.CONCLUSIONS The CRAB strain from the case 1 is brought from community,which differs in the evolution branch with the strains from the cases of hospi-tal-associated CRAB infection.The transmission of CRAB leading to the incident of suspected hospital-associated infection outbreak is multiple sources.It is an effective measure to early identify the outbreak of hospital-associated infection and take targeted prevention measures so as to control the outbreak of hospital-associated infection.

OBJECTIVE To conduct the epidemiological survey for 4 patients with carbapenem-resistant Klebsiella pneumoniae(CRAB)that gathered to emerge in intensive care unit(ICU)within a short period so as to find out the sources of suspected CRAB.METHODS The clinical data and epidemiological history of the 4 patients with hospital-associated infection with CRAB who were treated in Shandong Provincial Hospital Affiliated to Shandong First Medical University from Mar.31,2023 to Apr.7,2023 were retrospectively analyzed.The whole genome second-generation sequencing was carried out for the CRAB strains isolated from the patients and their surround-ings,and the sources of the CRAB strains were traced by means of the whole genome sequencing.RESULTS The result of the epidemiological survey showed that the case 1,2,3 and 4 were close in spatial position and had the similar drug resistance spectrum,and there might be cross infection.With the combination of the result of genome analysis,it was found that the strain from the case 1 was brought from the community and was the evolutionary branch different from other CRAB strains causing nosocomial infection;the strains from the case 2,3 and 4 had the close original sources with the CRAB strains from surroundings.CONCLUSIONS The CRAB strain from the case 1 is brought from community,which differs in the evolution branch with the strains from the cases of hospi-tal-associated CRAB infection.The transmission of CRAB leading to the incident of suspected hospital-associated infection outbreak is multiple sources.It is an effective measure to early identify the outbreak of hospital-associated infection and take targeted prevention measures so as to control the outbreak of hospital-associated infection.

Ulcerative colitis(UC)is a common chronic non-specific intestinal inflammatory disease clinically.Its pathogenesis is complex,prolonged the disease course,and it is frequent clinical recurrence.In recent years,the incidence of UC has shown an upward trend and tends to younger onset,which seriously affects the quality of life of patients and increases the social medical burden.Mitogen-activated protein kinase(MAPK)plays an important role in the occurrence and development of UC,participating in inflammatory response,oxidative stress,autophagy,apoptosis,pyroptosis,and intestinal barrier.MAPK may be a new target for the treatment of UC.Traditional Chinese medicine has a long history and remarkable curative effect in treating UC,with the advantages of multiple pathways,multiple targets,and multiple components.In recent years,many studies have shown that single Chinese herbal medicines and compound prescriptions can mediate MAPK signaling pathway to inhibit inflammatory response and oxidative stress,regulate autophagy,inhibit apoptosis and pyroptosis,repair the intestinal barrier,and treat UC in many aspects.This article reviews the MAPK signaling pathway,the mechanism by which the MAPK signaling pathway regulates UC,and the research progress of traditional Chinese medicine in treating UC based on the MAPK signaling pathway,hoping to provide theoretical support for the treatment of UC by traditional Chinese medicine and new ideas and references for the research of related new drugs.

Objective To investigate the relationship between ferroptosis-related genes[heme oxygenase-1(HO-1),solute carrier family 7 member 11(SLC7A11),and long-chain acyl-coenzyme A synthetase 4(ACSL4)]expression and cardiovascular events in patients with chronic renal failure(CRF),aiming to pro-vide clinical insights for risk assessment of cardiovascular complications.Methods A total of 124 CRF pa-tients(CRF group)admitted to the hospital from January 2020 to January 2023 and 124 healthy people(con-trol group)who underwent physical examination in the hospital during the same period were selected as the research objects.The levels of ferroptosis-related gene HO-1,SLC7A11 and ACSL4 were detected and com-pared between the two groups.CRF patients were followed up for 12 months after discharge,and they were di-vided into concurrent group(n=56)and non-concurrent group(n=66)according to the occurrence of cardio-vascular events.Spearman correlation analysis was used to investigate the correlation between iron death relat-ed genes HO-1,SLC7A11,ACSL4 and cardiovascular events in CRF patients.Univariate and multivariate Lo-gistic regression analysis were used to investigate the influencing factors of cardiovascular events in CRF pa-tients.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of iron death re-lated genes HO-1,SLC7A11,and ACSL4 for cardiovascular events in CRF patients.Results Serum HO-1 and SLC7A11 levels in CRF group were lower than those in control group(P＜0.05),and the ACSL4 level was higher than that in control group(P＜0.05).The serum HO-1 and SLC7A11 levels in concurrent group were lower than those in non-concurrent group(all P＜0.05),and the serum ACSL4 level was higher than that in non-concurrent group(P＜0.05).Ferroptosis-related genes HO-1 and SLC7A11 were negatively correlated with cardiovascular events(r=-0.708,—0.721,P＜0.05),while ACSL4 was positively correlated with car-diovascular events(r=0.699,P＜0.05).High serum ACSL4 expression and high cTnT level were risk fac-tors for cardiovascular events in CRF patients(P＜0.05),and high hemoglobin level and high serum HO-1 and SLC7A11 expression were protective factors for cardiovascular events in CRF patients(P＜0.05).ROC curve analysis results showed that the area under the curve(AUC)of serum HO-1,SLC7A11,ACSL4,hemo-globin and cTnT alone for predicting cardiovascular events in CRF patients were 0.787,0.735,0.773,0.651 and 0.782,respectively.The AUC of the combined prediction of ferroptosis-related genes was 0.837,and the AUC of the combined application of five factors was 0.880.According to Delong's test,the AUC of both com-bined application models was significantly higher than those of individual application(P＜0.05).The nomo-gram model showed that all ferroptosis-related genes were positively expressed(HO-1＜1.5,SLC7A11＜1.15,ACSL4≥2.75),and the risk of developing concurrent cardiovascular events in CRF patients could reach over 75％.Conclusion The expression of serum HO-1 and SLC7A11 in CRF patients is decreased,and the ACSL4 expression is increased,which is closely related to cardiovascular events,and the combined detection of three indexes has high value in predicting cardiovascular events in CRF patients.

Objective To establish a 13C isotope-labeled hyperinsulinemic-euglycemic clamp model for the assessment of insulin sensitivity in mice.Methods The mouse model of insulin resistance was established by high-fat diet feeding.The phosphorylation level of downstream insulin signaling protein,Protein Kinase,also known as Akt was assessed.Glucose metabolism was evaluated using glucose tolerance test,insulin tolerance test,and pyruvate tol-erance tests and the area under the curve of the respective testes over 2 hours was calculated to quantify overall blood glucose level.Mice underwent jugular vein catheterization surgery,and a 13C isotope-labeled hyperinsulinemic-eugly-cemic clamp experiment was conducted to monitor blood glucose levels and to calculate the glucose infusion rate(GIR).Tail vein serum was collected for liquid chromatography-tandem mass spectrometry(LC-MS/MS)analysis to determine glucose disposal rate(GDR)and hepatic gluconeogenesis rate(HGP).Following the inhibition of glycog-enolysis in mice,Akt phosphorylation level was measured to evaluate insulin signaling.The clamp test was repeated to calculate GIR,and tail vein blood serum was analyzed by LC-MS/MS to determine GDR and HGP.Results After one week of high-fat diet feeding,mice exhibited significantly elevated blood glucose level(P＜0.001)accompanied by reduced p-Akt level in liver and muscle.Glucose tolerance tests,insulin tolerance tests and pyruvate tolerance test demonstrated a significant increase in blood glucose level(P＜0.05)and a higher area under curves(AUC)(P＜0.001)in high-fat-fed mice.During the 13C-labeled hyperinsulinemic-euglycemic clamp experiment,after the blood glucose levels were stable,the GIR of high-fat-fed mice was significantly reduced(P＜0.001),GDR was de-creased(P＜0.0001)and hepatic gluconeogenesis rate was increased(P＜0.01).After pharmacological inhibition of glycogenolysis,mice showed elevated blood glucose level(P＜0.001)and further reductions in p-Akt level in liver and muscle.The 13C-labeled clamp experiment revealed that in the treated group,the GIR decreased(P＜0.01)while GDR was reduced(P＜0.000 1)and HGP increased(P＜0.01).Conclusions An improved hyperinsulinemic-eugly-cemic clamp model was developed to assess insulin sensitivity in mice.

Cancer incidence in China has been rising steadily,with a particularly heavy burden from several high-prevalence malignancies.Medical examination for cancer plays a critical role in the early detection of cancer,precancerous lesions,and precursor conditions,thereby facilitating timely diagnosis and intervention.Such examination also addresses the growing demand for person-alized cancer screening services among diverse population groups.The development of evidence-based,context-specific cancer screening guidelines is essential to enhance the standardization,quality,and equity of preventive screening practices across the country,ultimately improving out-comes in early cancer detection and treatment.Guided by the Department of Medical Emergency Response of the National Health Commission,the Guidelines for Medical Examination for Cancer in Health Examination Agency(2025 Edition)were developed under the leadership of the National Cancer Center.A multidisciplinary panel of experts formulated the guidelines in accordance with the principles and methodology of the World Health Organization Handbook for Guideline Deve-lopment.The guidelines provide evidence-based recommendations on key clinical domains:target cancers and populations,overall screening workflow,screening protocols,diagnostic technolo-gies,result interpretation,follow-up procedures,and quality control.The primary objective is to standardize cancer screening practices in health examination agency and strengthen China's ca-pacity for prevention and control of high-burden cancers.

Objective:To explore the impact of the stimulator of interferon genes(STING)-interferon regulatory factor 3(IRF3)pathway on the senescence of rapid pacing HL-1 myocytes.Methods:HL-1 cells were divided into five groups: the control group(HL-1 cells without any treatment), pacing group(HL-1 cells paced for 48 hours), STING siRNA group(HL-1 cells paced for 48 hours and transfected with STING siRNA), NC siRNA group(HL-1 cells paced for 48 hours and transfected with NC siRNA), and H151 inhibitor group(HL-1 cells paced for 48 hours with the addition of 1 μmol/L STING inhibitor H151). Mitochondrial membrane potential was assessed in control and pacing group cells, and mitochondrial MitoTracker and TFAM co-localization staining was performed on these cells.Cellular senescence was evaluated using β-galactosidase staining in each group, and the positive rate of cellular senescence was observed and calculated.Western blotting was employed to detect the expression levels of STING, IRF3, P-IRF3, P16, P21, and P53 proteins in all groups.Immunofluorescence was utilized to examine the expression distribution of STING and P21 across the various groups.ELISA was performed to measure the concentrations of interleukin(IL)-1β, IL-6, and IL-8 in the cell supernatants from each group as part of the senescence-associated secretory phenotype(SASP).Results:Compared with the control group, the ratio of mitochondrial JC-1 multimer to monomer was significantly decreased in the pacing group( t=16.42, P<0.05), the co-localization of mitochondrial MitoTracker and TFAM in the cells was significantly weakened, the proportion of cells with positive cellular senescence-associated β-galactosidase staining significantly increased in the pacing group, the expression levels of STING, P-IRF3/IRF3, P16, P21, and P53 proteins were significantly elevated in the pacing group, and the concentrations of IL-1β, IL-6, and IL-8 in the cell supernatants were markedly increased.Compared with the pacing group, the proportion of cells with positive cellular senescence-associated β-galactosidase staining decreased in the STING siRNA group and H151 inhibitor group ( F= 18.13, P<0.05), the expression levels of STING, P-IRF3/IRF3, P16, P21, and P53 were reduced in the STING siRNA group and H151 inhibitor group ( F=16.84, 26.56, 74.70, 31.80, 31.23, all P<0.05), and the concentrations of IL-1β, IL-6, and IL-8 in the cell supernatants decreased( F=197.80、1 339.00、1 308.00, all P<0.001). Conclusions:Rapid pacing of HL-1 cells can promote mtDNA release into the cytoplasm, activate the STING-IRF3 pathway, accelerate cellular senescence, and enhance the secretion of SASP.Inhibiting the expression of STING can delay the senescence induced by the rapid pacing of HL-1 cells and reduce SASP secretion.

Dry eye disease (DED) is a prevalent and intractable ocular disease induced by a variety of causes. Elevated sphingomyelin (SM) levels and pro-inflammatory cytokines were detected on the ocular surface of DED patients, particularly in the meibomian glands. Sphingomyelin synthase 2 (SMS2), one of the proteins involved in SM synthesis, would light a novel way of developing a DED therapy strategy. Herein, we report the design and optimization of a series of novel thiophene carboxamide derivatives to afford 14l with an improved highly potent inhibitory activity on SM synthesis (IC_{50, SMS2} = 28 nmol/L). Moreover, 14l exhibited a notable protective effect of anti-inflammation and anti-apoptosis on human corneal epithelial cells (HCEC) under TNF-α-hyperosmotic stress conditions in vitro, with an acceptable ocular specific distribution (corneas and meibomian glands) and pharmacokinetics (PK) profiles (t _{1/2, cornea} = 1.11 h; t _{1/2, meibomian glands} = 4.32 h) in rats. Furthermore, 14l alleviated the dry eye symptoms including corneal fluorescein staining scores and tear secretion in a dose-dependent manner in mice. Mechanically, 14l reduced the mRNA expression of Tnf-α, Il-1β and Mmp-9 in corneas, as well as the proportion of very long chain SM in meibomian glands. Our findings provide a new strategy for DED therapy based on selective SMS2 inhibitors.