Objective:To evaluate the predictive performance of machine learning methods for predicting the risk of death in traumatic hemorrhage, and address the low prediction accuracy of traditional trauma scores, provide a reference for developing a more robust prediction method for severe trauma patients.Methods:Clinical data of severe trauma patients from the National Trauma Medical Center between April 1, 2023, and March 31, 2024 were collected. ElasticNet, Recursive Feature Elimination, and Mutual Information-based feature selection methods were used to screen variables and compared with traditional hypothesis testing methods. Built the prediction models for mortality risk in traumatic hemorrhage using Logistic Regression, ElasticNet, and Support Vector Machine (SVM) and compared the predictive performance.Results:The study included 5,601 trauma patients, the results of the variable screening and importance ranking were consistent with three feature selection methods. The classification accuracy and AUC values for the three models were as follows: Overall accuracy was 83.2%, survival accuracy was 84.0%, death accuracy was 76.3%, and an AUC was 0.86 in logistic regression; Overall accuracy was 78.9%, survival accuracy was 78.5%, death accuracy was 81.7%, and an AUC was 0.88 in ElasticNet; Overall accuracy was 84.7%, survival accuracy was 86.1%, death accuracy was 72.4%, and an AUC was 0.86 in SVM. The prediction performance of three models is quite little.Conclusion:Machine learning methods can effectively improve the prediction of death risk for traumatic hemorrhage,and has wide applications.

AIM:To elucidate the intervention and mechanism of 4'-hydroxychalcone(4-HC)in colitis mice through the regulation of Th17/Treg homeostasis.METHODS:Using a dextran sodium sulfate(DSS)-induced colitis model in mice,we meticulously observed the pathological characteristics of colon tissue via HE staining.Additionally,we employed immunohistochemical analysis and Western blot techniques to assess the expression levels of proteins associated with the JAK/STAT signaling pathway,as well as the specific content of tight junction proteins such as ZO-1 and occludin.The differentiation of Th17 and Treg cells was analyzed through flow cytometry.RESULTS:Compared to the normal group,the DSS group exhibited a consistent decline in body weight,coupled with symptoms of diarrhea and hematochezia,an increase in the DAI score,and a notable reduction in colon length.In contrast,the body weight of the 4-HC group dis-played an upward trend following an initial decrease,with improvements in diarrhea and hematochezia symptoms,a reduc-tion in the DAI score,and a restoration of colon length relative to the model group.The integrity of colon tissue in the 4-HC group was significantly better than that in the DSS group,evidenced by a marked increase in the number of goblet cells and an enhancement in crypt integrity,while the average histology score showed a decrease.Western blot analysis re-vealed substantial increase in ZO-1 and occludin expression after 4-HC treatment.Flow cytometry results indicated a dra-matic decrease in the differentiation rate of Th17 cells in spleen lymphocytes and mesenteric lymph nodes,while the dif-ferentiation rate of Treg cells was significantly elevated.Immunohistochemical and Western blot analyses demonstrated that 4-HC markedly reduced the phosphorylation of STAT1 and STAT3,while up-regulating the phosphorylation of STAT6,suggesting that 4-HC modulates CD4+T cell activity through the JAK-STAT pathway.CONCLUSION:The 4-HC may enhance the course of DSS-induced colitis in mice,alleviate colonic tissue damage,and modulate the balance be-tween Th17 and Treg cells,potentially involving the JAK/STAT signaling pathway.

Objective To analyze the drug resistance characteristics,molecular typing,and biological properties of carbapenem-resist-ant Klebsiella pneumoniae(CRKP).Methods A retrospective analysis was conducted on 31 non-repetitive CRKP strains collected clinically from April 2019 to May 2021 at the Third People's Hospital of Nantong.The Vitek 2 Compact microbial analysis system was used for bacterial identification and in vitro drug susceptibility analysis.The broth dilution method was used to determine the minimum inhibitory concentration(MIC)of polymyxin B.The disk diffusion testing was performed to supplement the susceptibility of five com-monly used antibiotics:ertapenem,cefotaxime,cefoxitin,cefoperazone-sulbactam,and tigecycline.The carbapenemase-resistance phenotype of CRKP strains was initially determined by a combined assay of modified carbapenem inactivation method(mCIM)and ED-TA-carbapenem inactivation method(eCIM).Certain carbapenemase resistance genes(blaKPC,blaNDM,blaIMP,blaVIM,and blaOXA-48),AmpC enzyme genes(blaDHA,bla ACC,blaCIT,blaEBC,blaMOX,and blaFOX),extended-spectrum β-lactamases(ESBLs)genes(blaSHv,blaTEM,and blaCTX-M),and nine virulence genes were amplified by PCR and subsequently verified by sequencing.The stringing test was used to screen for hypermucoviscous phenotype strains.The growth curves in vitro and biofilm formation assays,and multilocus se-quence typing(MLST)were performed on 31 isolates.Outer-membrane proteins were extracted and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)to evaluate the expressions of OmpK35 and OmpK36.Results All the 31 isolates were resistant to ampicillin/sulbactam,ampicillin,aztreonam,cefazolin,ceftriaxone,cefotaxime,cefuroxime,ciprofloxacin,pip-eracillin,piperacillin/tazobactam,meropenem,and ertapenem with resistance rate of 100％.The resistance to polymyxin B was ob-served in 32.26％,whereas tigecycline retained 100％susceptibility.In terms of MLST,three sequence types(STs)were identified,with ST15 being the most prevalent,accounting for 61.29％(19/31)of the isolates.All strains produced serine carbapenemase,and only blaKPC-2 was detected among carbapenem resistance genes.The virulence genes fimH and entB were present in all strains(100％,31/31),while the detection rate of mrkD was 80.64％(25/31).Some strains carried virulence genes such as rmpA,rmpA2,and other virulence genes,whereas magA gene was not detected in any isolate.The carriage rates of rmpA2,iutA,and mrkD were higher in ST11 strains than in ST15 strains.The string test was positive in 38.71％of the strains.The growth test showed that there was no significant difference observed in the growth curves among all strains in vitro,and all were able to form biofilms with varying degrees.All ST11 strains exhibited OmpK36 protein alterations,while OmpK35 protein was intact in the 31 strains.Conclusion CRKP strains in this hospital showed high drug-resistance rate,and ST15 was the predominant sequence type.All the isolates carried blaKPC-2 and virulence genes.Enhanced molecular surveillance and strengthened prevention and control measures of CRKP infection are urgently needed.

Objective To analyze the drug resistance characteristics,molecular typing,and biological properties of carbapenem-resist-ant Klebsiella pneumoniae(CRKP).Methods A retrospective analysis was conducted on 31 non-repetitive CRKP strains collected clinically from April 2019 to May 2021 at the Third People's Hospital of Nantong.The Vitek 2 Compact microbial analysis system was used for bacterial identification and in vitro drug susceptibility analysis.The broth dilution method was used to determine the minimum inhibitory concentration(MIC)of polymyxin B.The disk diffusion testing was performed to supplement the susceptibility of five com-monly used antibiotics:ertapenem,cefotaxime,cefoxitin,cefoperazone-sulbactam,and tigecycline.The carbapenemase-resistance phenotype of CRKP strains was initially determined by a combined assay of modified carbapenem inactivation method(mCIM)and ED-TA-carbapenem inactivation method(eCIM).Certain carbapenemase resistance genes(blaKPC,blaNDM,blaIMP,blaVIM,and blaOXA-48),AmpC enzyme genes(blaDHA,bla ACC,blaCIT,blaEBC,blaMOX,and blaFOX),extended-spectrum β-lactamases(ESBLs)genes(blaSHv,blaTEM,and blaCTX-M),and nine virulence genes were amplified by PCR and subsequently verified by sequencing.The stringing test was used to screen for hypermucoviscous phenotype strains.The growth curves in vitro and biofilm formation assays,and multilocus se-quence typing(MLST)were performed on 31 isolates.Outer-membrane proteins were extracted and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)to evaluate the expressions of OmpK35 and OmpK36.Results All the 31 isolates were resistant to ampicillin/sulbactam,ampicillin,aztreonam,cefazolin,ceftriaxone,cefotaxime,cefuroxime,ciprofloxacin,pip-eracillin,piperacillin/tazobactam,meropenem,and ertapenem with resistance rate of 100％.The resistance to polymyxin B was ob-served in 32.26％,whereas tigecycline retained 100％susceptibility.In terms of MLST,three sequence types(STs)were identified,with ST15 being the most prevalent,accounting for 61.29％(19/31)of the isolates.All strains produced serine carbapenemase,and only blaKPC-2 was detected among carbapenem resistance genes.The virulence genes fimH and entB were present in all strains(100％,31/31),while the detection rate of mrkD was 80.64％(25/31).Some strains carried virulence genes such as rmpA,rmpA2,and other virulence genes,whereas magA gene was not detected in any isolate.The carriage rates of rmpA2,iutA,and mrkD were higher in ST11 strains than in ST15 strains.The string test was positive in 38.71％of the strains.The growth test showed that there was no significant difference observed in the growth curves among all strains in vitro,and all were able to form biofilms with varying degrees.All ST11 strains exhibited OmpK36 protein alterations,while OmpK35 protein was intact in the 31 strains.Conclusion CRKP strains in this hospital showed high drug-resistance rate,and ST15 was the predominant sequence type.All the isolates carried blaKPC-2 and virulence genes.Enhanced molecular surveillance and strengthened prevention and control measures of CRKP infection are urgently needed.

AIM:To elucidate the intervention and mechanism of 4'-hydroxychalcone(4-HC)in colitis mice through the regulation of Th17/Treg homeostasis.METHODS:Using a dextran sodium sulfate(DSS)-induced colitis model in mice,we meticulously observed the pathological characteristics of colon tissue via HE staining.Additionally,we employed immunohistochemical analysis and Western blot techniques to assess the expression levels of proteins associated with the JAK/STAT signaling pathway,as well as the specific content of tight junction proteins such as ZO-1 and occludin.The differentiation of Th17 and Treg cells was analyzed through flow cytometry.RESULTS:Compared to the normal group,the DSS group exhibited a consistent decline in body weight,coupled with symptoms of diarrhea and hematochezia,an increase in the DAI score,and a notable reduction in colon length.In contrast,the body weight of the 4-HC group dis-played an upward trend following an initial decrease,with improvements in diarrhea and hematochezia symptoms,a reduc-tion in the DAI score,and a restoration of colon length relative to the model group.The integrity of colon tissue in the 4-HC group was significantly better than that in the DSS group,evidenced by a marked increase in the number of goblet cells and an enhancement in crypt integrity,while the average histology score showed a decrease.Western blot analysis re-vealed substantial increase in ZO-1 and occludin expression after 4-HC treatment.Flow cytometry results indicated a dra-matic decrease in the differentiation rate of Th17 cells in spleen lymphocytes and mesenteric lymph nodes,while the dif-ferentiation rate of Treg cells was significantly elevated.Immunohistochemical and Western blot analyses demonstrated that 4-HC markedly reduced the phosphorylation of STAT1 and STAT3,while up-regulating the phosphorylation of STAT6,suggesting that 4-HC modulates CD4+T cell activity through the JAK-STAT pathway.CONCLUSION:The 4-HC may enhance the course of DSS-induced colitis in mice,alleviate colonic tissue damage,and modulate the balance be-tween Th17 and Treg cells,potentially involving the JAK/STAT signaling pathway.

In order to develop a positive quality control products for the detection of porcine repro-ductive and respiratory syndrome virus(PRRSV)nucleic acid by real-time fluorescent quantitative PCR(RT-qPCR),the positive quality control products of PRRSV-1 and PRRSV-2 M genes were prepared using armored RNA technology of MS2 phage.PRRSV-1 and PRRSV-2 M genes were amplified,purified and recovered,and ligated into pET28b vector containing MS2 mature enzyme protein gene and capsid protein.After transformed into BL21(DE3),the gene products were in-duced by IPTG and purified by PEG6000 precipitation method to prepare the armored RNA virus-like particles(AR-PRRSV)containing PRRSV M gene.Following the performance evaluation,as the positive quality control products of PRRSV-1 and PRRSV-2 M genes,AR-PRRSV1M and AR-PRRSV2M were calculated using YY/T 1652-2019 standard.Results showed that it had a good u-niformity,stable storage for the armored virus-like particles at-20,4,25 ℃ for 60 d,and 37 ℃ for 30 d.The prepared armored virus-like particles AR-PRRSV1M and AR-PRRSV2M were deter-mined by digital quantitative PCR(ddPCR)after preliminary quantification by RT-qPCR.The 104 copies/μL of AR-PRRSV1M and AR-PRRSV2M ddPCR fixation was(1.33+0.50)× 104 cop-ies/μL.The above results indicates that the AR-PRRSVM can be used as the quality control of the whole detection process(nucleic acid extraction,reverse transcription and RT-qPCR).

AIM:To investigate the potential of atractylenolide Ⅲ(AⅢ)in mitigating dextran sulfate sodium(DSS)-induced injury in a mouse model of chronic inflammatory bowel disease(IBD),and to explore the mechanisms in-volved,particularly the modulation of signal transducer and activator of transcription 3(STAT3)signaling,which plays a crucial role in the homeostasis of T helper 17(Th17)and regulatory T(Treg)cells.METHODS:A mouse model of DSS-induced chronic IBD was established,and the mice were divided into 4 groups:control,model(DSS),high-dose(50 mg/kg)AⅢ,and low-dose(30 mg/kg)AⅢ.The disease activity index(DAI)was utilized to assess disease severity.Histo-pathological damage in the colons of IBD mice was evaluated by hematoxylin-eosin(HE)staining.The protein levels of phosphorylated STAT3,occludin and zonula occludens-1(ZO-1)were analyzed using immunohistochemical staining and Western blot.Flow cytometry was employed to examine the differentiation of splenic lymphocytes into Th17/Treg cells.RESULTS:Both DAI assessments and HE staining indicated that AⅢ significantly alleviated inflammatory injury in mice with DSS-induced chronic IBD.Immunohistochemical analysis demonstrated that AⅢ enhanced the expression of ZO-1 and occludin in colonic tissues.Flow cytometry results revealed that AⅢ helped maintain the balance between splenic Th17 and Treg cells.Furthermore,immunohistochemical staining and Western blot showed that AⅢ inhibited the phos-phorylation of STAT3.CONCLUSION:Treatment with AⅢ effectively reduced inflammatory injury in a mouse model of chronic IBD by preserving Th17/Treg homeostasis through the inhibition of STAT3 phosphorylation.As a natural com-pound,AⅢ exhibits significant therapeutic potential for the treatment of chronic IBD.

AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Objective: To understand the current situation of health rooms in primary and secondary schools in Jilin Province, so as to provide the data support for scientific decision making. Methods: From April to July 2023, 512 primary and secondary schools and 1 432 school doctors and health care teachers were selected through convenience sampling method in Jilin Province to conduct an electronic questionnaire survey, including the basic information of the school, the situation of health rooms, personnel setting, and the development of school health work. Results: Among the 512 schools, only 6.4% of the 299 schools that should have clinics had medical institution practice licenses. The compliance rate of clinic area was 16.6%, and the compliance rate of health room area was 75.0%. About 92.1% of the middle schools and 90.6% of the primary schools identified the reporters of infectious diseases, and 90.9% of the primary schools and 85.5% of the secondary schools filed files for students. Totally 73.5% of the staff in the health room were teachers, and only 17.9% were health professionals. Nearly 70.1% of school doctors or health care teachers were engaged in part time jobs, and 60.9% engaged in school health for ≤5 years. In terms of the content in urgent need of training and improvement, the top five were knowledge about first aid (79.7%)，infectious disease prevention and treatment( 73.3 %), health education (64.0%), common disease diagnosis (60.1%) and psychological counseling (53.6%). Conclusions Health care institutions, equipment and facilities in primary and secondary schools in Jilin Province are inadequate, and the construction of school doctors and health care teachers is in need of improvement. It should pay more attention to school health and work together to optimize the team of school doctors and health care teachers.