ObjectiveTo investigate the mechanisms by which Mahuang Lianqiao Chixiaodoutang （MLC） improves obesity-type polycystic ovary syndrome （PCOS） through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodsThirty-six female Sprague-Dawley （SD） rats were randomly divided into a blank control group （Con） and an obesity-type PCOS model preparation group. The model was induced by gavage with letrozole （1 mg·kg^-1） combined with a high-fat diet （HFD）. After model establishment， the obesity-type PCOS model preparation group was further divided into the model group （Mod， normal saline）， metformin group （Met， 0.3 g·kg^-1）， low-dose MLC group （MLC-L， 4.3 g·kg^-1）， medium-dose MLC group （MLC-M， 8.6 g·kg^-1）， and high-dose MLC group （MLC-H， 17.2 g·kg^-1）. Active components of MLC and targets of obesity-type PCOS were screened from databases， a protein-protein interaction （PPI） network was constructed， and gene ontology（GO）and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis was performed. The gut microbiota structure was analyzed based on 16S rRNA sequencing and correlated with network pharmacology pathways. Body weight and estrous cycle were dynamically monitored. Ovarian morphology was observed by hematoxylin-eosin （HE） staining. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）. Enzyme-linked immunosorbent assay （ELISA） was used to detect levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， anti-Müllerian hormone （AMH）， testosterone （T）， and estradiol （E₂）. Western blot was used to detect the protein expression levels of phosphorylated PI3K/PI3K （p-PI3K/PI3K）， phosphorylated Akt/Akt （p-Akt/Akt）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax）. ResultsNetwork pharmacology screening identified 124 active components of MLC and 408 overlapping targets between the herbal formula and the disease. Core targets such as Akt1 and Bcl-2 were revealed. As indicated by 16S rRNA sequencing， the abundances of Lachnospiraceae， Lachnoclostridium， and Dorea were increased in the MLC groups （P<0.05）， while the abundance of Veillonella was decreased （P<0.05）. KEGG correlation analysis integrating network pharmacology and gut microbiota data showed significant enrichment of the PI3K/Akt signaling pathway. Animal experiments showed that， compared with the Mod group， body weight decreased to normal levels in the Met， MLC-M， and MLC-H groups. The estrous cycle became regular. The number of corpora lutea increased and cystic follicles decreased. Serum levels of T， FSH， and LH/FSH were reduced （P<0.05， P<0.01）， while the E₂ level was increased （P<0.01）. Ovarian cell apoptosis was reduced （P<0.01）， and the protein expression levels of p-PI3K/PI3K， p-Akt/Akt， and Bcl-2 in ovarian tissue were significantly increased， whereas Bax protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionMLC can regulate gut microbiota structure， effectively improve ovarian pathology in rats with obesity-type PCOS， and inhibit ovarian granulosa cell apoptosis. The mechanism may be associated with upregulation of the PI3K/Akt signaling pathway.

ObjectiveTo investigate the mechanisms by which Mahuang Lianqiao Chixiaodoutang （MLC） improves obesity-type polycystic ovary syndrome （PCOS） through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodsThirty-six female Sprague-Dawley （SD） rats were randomly divided into a blank control group （Con） and an obesity-type PCOS model preparation group. The model was induced by gavage with letrozole （1 mg·kg^-1） combined with a high-fat diet （HFD）. After model establishment， the obesity-type PCOS model preparation group was further divided into the model group （Mod， normal saline）， metformin group （Met， 0.3 g·kg^-1）， low-dose MLC group （MLC-L， 4.3 g·kg^-1）， medium-dose MLC group （MLC-M， 8.6 g·kg^-1）， and high-dose MLC group （MLC-H， 17.2 g·kg^-1）. Active components of MLC and targets of obesity-type PCOS were screened from databases， a protein-protein interaction （PPI） network was constructed， and gene ontology（GO）and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis was performed. The gut microbiota structure was analyzed based on 16S rRNA sequencing and correlated with network pharmacology pathways. Body weight and estrous cycle were dynamically monitored. Ovarian morphology was observed by hematoxylin-eosin （HE） staining. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）. Enzyme-linked immunosorbent assay （ELISA） was used to detect levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， anti-Müllerian hormone （AMH）， testosterone （T）， and estradiol （E₂）. Western blot was used to detect the protein expression levels of phosphorylated PI3K/PI3K （p-PI3K/PI3K）， phosphorylated Akt/Akt （p-Akt/Akt）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax）. ResultsNetwork pharmacology screening identified 124 active components of MLC and 408 overlapping targets between the herbal formula and the disease. Core targets such as Akt1 and Bcl-2 were revealed. As indicated by 16S rRNA sequencing， the abundances of Lachnospiraceae， Lachnoclostridium， and Dorea were increased in the MLC groups （P<0.05）， while the abundance of Veillonella was decreased （P<0.05）. KEGG correlation analysis integrating network pharmacology and gut microbiota data showed significant enrichment of the PI3K/Akt signaling pathway. Animal experiments showed that， compared with the Mod group， body weight decreased to normal levels in the Met， MLC-M， and MLC-H groups. The estrous cycle became regular. The number of corpora lutea increased and cystic follicles decreased. Serum levels of T， FSH， and LH/FSH were reduced （P<0.05， P<0.01）， while the E₂ level was increased （P<0.01）. Ovarian cell apoptosis was reduced （P<0.01）， and the protein expression levels of p-PI3K/PI3K， p-Akt/Akt， and Bcl-2 in ovarian tissue were significantly increased， whereas Bax protein expression was significantly decreased （P<0.05， P<0.01）. ConclusionMLC can regulate gut microbiota structure， effectively improve ovarian pathology in rats with obesity-type PCOS， and inhibit ovarian granulosa cell apoptosis. The mechanism may be associated with upregulation of the PI3K/Akt signaling pathway.

ObjectiveTo investigate the role of interleukin （IL）-17A in acute inhalational pneumonia induced by the highly drug-resistant and hypervirulent Staphylococcus aureus strain USA300-R in mice. MethodsAn acute inhalational pneumonia model was established in mice using an aerosolized pulmonary delivery technique. RNA sequencing （RNA-seq） and enzyme-linked immunosorbent assay （ELISA） were employed to examine the expression dynamics of Il17a mRNA and IL-17A protein， respectively， in the lungs of infected mice. Il17a knockout （Il17a^-/-） mice were generated using CRISPR/Cas9 gene editing technology. The survival rate， body weight， bacterial load in lung tissue， and histopathological changes were compared between Il17a^-/- and wild-type （WT） mice following inhalational infection with USA300-R. Results12 hours after USA300-R infection， compared to pre-infection， the expression level of Il17a mRNA in lung tissue and the level of IL-17A protein in bronchoalveolar lavage fluid （BALF） increased by approximately 50-fold （P<0.01） and 6-fold （P<0.001）， respectively. Compared to WT mice， Il17a^-/- mice exhibited approximately 10-fold higher bacterial loads in lung tissue at both 12 and 24 hours post-infection （P<0.001， P<0.05）. However， they showed significantly attenuated lung histopathological injury， reduced alveolar wall thickening， markedly decreased neutrophil infiltration， and an approximately 50% improvement in survival rate （P<0.05）. ConclusionIn acute Staphylococcus aureus USA300-R inhalational pneumonia， IL-17A contributes to bacterial clearance by recruiting neutrophils； however， excessive neutrophil infiltration exacerbates pulmonary inflammation and injury， reduces survival rates， and represents a potential therapeutic target.

Objective:To investigate the effect of graded rehabilitation intervention on cardiac function,self-man-agement ability,exercise tolerance,sleep quality and quality of life in patients with chronic heart failure(CHF).Methods:This randomized controlled study enrolled 125 patients with CHF admitted to Lu'an People's Hospital be-tween April 2020 and October 2023.Patients were divided into control group(n=62)and intervention group(n=63).Participants in the control group received routine rehabilitation intervention,while those in the intervention group received additional graded rehabilitation intervention.Both groups were intervened for 3 months.Cardiac function indexes,6min walking distance(6MWD),metabolic equivalent(METs),scores of exercise of self-care a-gency(ESCA),Pittsburgh sleep quality index(PSQI)and the MOS item short from health survey(SF-36),and in-cidence of adverse events during the intervention were compared between the two groups.Results:Compared to pa-tients in the control group,those in intervention group had significant lower left ventricular end-diastolic volume(LVEDV)[(108.19±2.96)ml vs.(129.51±3.34)ml],left ventricular end-systolic volume(LVESV)[(50.68±2.28)ml vs.(67.38±2.68)ml],score of Pittsburgh sleep quality index(PSQI)[(8.30±1.87)points vs.(13.99±2.06)points](P＜0.001 all),and significant higher left ventricular ejection fraction(LVEF)[(58.17±2.08)％vs.(50.70±1.63)％],ESCA score[(148.29±19.48)points vs.(124.68±15.93)points],6MWD[(413.58±15.11)m vs.(372.22±14.62)m],METs[(6.88±0.64)kcal·kg-1·h-1 vs.(6.13±0.62)kcal·kg-1·h-1],total SF-36 score[(128.97±6.62)points vs.(104.35±5.46)points](P＜0.001 all).The inci-dence of adverse events in the intervention group was significantly lower than that in the control group(5.00％vs.18.33％,P=0.023).Conclusion:The graded rehabilitation intervention may improve the heart function,self-management ability,exercise tolerance,sleep quality and quality of life in CHF patients,and has high safety.

Objective:To investigate the effect of graded rehabilitation intervention on cardiac function,self-man-agement ability,exercise tolerance,sleep quality and quality of life in patients with chronic heart failure(CHF).Methods:This randomized controlled study enrolled 125 patients with CHF admitted to Lu'an People's Hospital be-tween April 2020 and October 2023.Patients were divided into control group(n=62)and intervention group(n=63).Participants in the control group received routine rehabilitation intervention,while those in the intervention group received additional graded rehabilitation intervention.Both groups were intervened for 3 months.Cardiac function indexes,6min walking distance(6MWD),metabolic equivalent(METs),scores of exercise of self-care a-gency(ESCA),Pittsburgh sleep quality index(PSQI)and the MOS item short from health survey(SF-36),and in-cidence of adverse events during the intervention were compared between the two groups.Results:Compared to pa-tients in the control group,those in intervention group had significant lower left ventricular end-diastolic volume(LVEDV)[(108.19±2.96)ml vs.(129.51±3.34)ml],left ventricular end-systolic volume(LVESV)[(50.68±2.28)ml vs.(67.38±2.68)ml],score of Pittsburgh sleep quality index(PSQI)[(8.30±1.87)points vs.(13.99±2.06)points](P＜0.001 all),and significant higher left ventricular ejection fraction(LVEF)[(58.17±2.08)％vs.(50.70±1.63)％],ESCA score[(148.29±19.48)points vs.(124.68±15.93)points],6MWD[(413.58±15.11)m vs.(372.22±14.62)m],METs[(6.88±0.64)kcal·kg-1·h-1 vs.(6.13±0.62)kcal·kg-1·h-1],total SF-36 score[(128.97±6.62)points vs.(104.35±5.46)points](P＜0.001 all).The inci-dence of adverse events in the intervention group was significantly lower than that in the control group(5.00％vs.18.33％,P=0.023).Conclusion:The graded rehabilitation intervention may improve the heart function,self-management ability,exercise tolerance,sleep quality and quality of life in CHF patients,and has high safety.

OBJECTIVE To develop a nomogram model for prediction of the risk of sepsis in the patients with acute liver failure(ALF)and validate its clinical value.METHODS Totally 228 patients with ALF who were treated in the Fifth Medical Center of Chinese PLA General Hospital from Jan.2009 to Mar.2023 were recruited as the re-search subjects and were grouped according to the occurrence of sepsis,the clinical characteristics and results of laboratory tests were collected from the patients.The subjects were brought into multivariate logistic regression a-nalysis after the primary screening with univariate regression analysis,the independent predictive factors were screened out,and the nomogram model was established.The accuracy,calibration accuracy and clinical practica-bility of the model were assessed by means of receiver operating characteristic(ROC)curves,calibration curves and decision curve analysis(DCA).RESULTS Among the 228 patients with ALF,159(69.74％)were diagnosed with sepsis.Totally 6 independent predictive factors were screened out by the multivariate analysis,including age[odd ratio(OR)=1.098,95％CI:1.030 to 1.220],aspartate transaminase(OR=0.998,95％CI:0.996 to 0.999),white blood cells counts(OR=1.037,95％CI:1.020 to 1.064),hemoglobulin(OR=0.981,95％CI:0.962 to 0.998),lactic acid(OR=1.187,95％CI:1.022 to 1.426)and mechanical ventilation(OR=3.463,95％CI:2.340 to 5.125).The area under the curve(AUC)of the nomogram model was 0.864(95％CI:0.807 to 0.921)in the training set,0.817(95％CI:0.717 to 0.918)in the validation set,remarkably better than that of sequential organ failure assessment(SOFA)scores[0.710(95％CI:0.625 to 0.795)and 0.647(95％CI:0.515 to 0.779)].Hosmer-Lemeshow test(P=0.512)and the calibrated curves showed that the predication probability of the model was highly consistent with the actual risk,and DC A indicated that the net clinical benefit was brought more from the model than from SOFA score.CONCLUSION The age,hemoglobulin,aspartate transaminase,white blood cells counts,lactic acid and mechanical ventilation are the independent predictive factors for the sepsis in the ALF patients.The nomogram model established based on the factors has high predictive efficiency and clini-cal application value.

OBJECTIVE To develop a nomogram model for prediction of the risk of sepsis in the patients with acute liver failure(ALF)and validate its clinical value.METHODS Totally 228 patients with ALF who were treated in the Fifth Medical Center of Chinese PLA General Hospital from Jan.2009 to Mar.2023 were recruited as the re-search subjects and were grouped according to the occurrence of sepsis,the clinical characteristics and results of laboratory tests were collected from the patients.The subjects were brought into multivariate logistic regression a-nalysis after the primary screening with univariate regression analysis,the independent predictive factors were screened out,and the nomogram model was established.The accuracy,calibration accuracy and clinical practica-bility of the model were assessed by means of receiver operating characteristic(ROC)curves,calibration curves and decision curve analysis(DCA).RESULTS Among the 228 patients with ALF,159(69.74％)were diagnosed with sepsis.Totally 6 independent predictive factors were screened out by the multivariate analysis,including age[odd ratio(OR)=1.098,95％CI:1.030 to 1.220],aspartate transaminase(OR=0.998,95％CI:0.996 to 0.999),white blood cells counts(OR=1.037,95％CI:1.020 to 1.064),hemoglobulin(OR=0.981,95％CI:0.962 to 0.998),lactic acid(OR=1.187,95％CI:1.022 to 1.426)and mechanical ventilation(OR=3.463,95％CI:2.340 to 5.125).The area under the curve(AUC)of the nomogram model was 0.864(95％CI:0.807 to 0.921)in the training set,0.817(95％CI:0.717 to 0.918)in the validation set,remarkably better than that of sequential organ failure assessment(SOFA)scores[0.710(95％CI:0.625 to 0.795)and 0.647(95％CI:0.515 to 0.779)].Hosmer-Lemeshow test(P=0.512)and the calibrated curves showed that the predication probability of the model was highly consistent with the actual risk,and DC A indicated that the net clinical benefit was brought more from the model than from SOFA score.CONCLUSION The age,hemoglobulin,aspartate transaminase,white blood cells counts,lactic acid and mechanical ventilation are the independent predictive factors for the sepsis in the ALF patients.The nomogram model established based on the factors has high predictive efficiency and clini-cal application value.

Objective To investigate the impact of temperature intervention on pain and catheterization outcomes in premature infants undergoing peripherally inserted central catheter(PICC)placement.Methods A total of 70 premature infants who underwent PICC placement and met the inclusion criteria were selected from the Neonatology Department of a tertiary children's hospital in Kunming,Yunnan,from January 2023 to December 2023.They were randomly divided into a control group(n=35)and an observation group(n=35)using a random number table.The control group underwent the standard PICC placement procedure;the observation group underwent temperature intervention measures based on the control group,which included using iodine tincture warmed to 37℃for disinfection,applying warm compresses to the catheterization site with heated hands of the operator,and using saline warmed to 37℃during catheter insertion and skin cleansing.Pain scores measured by the Premature Infant Pain Profile(PIPP),blood oxygen saturation,heart rate,success rates of single puncture and single catheter insertion,incidence of complications,and occurrence of hypothermia post-catheterization were compared between the two groups.Results The PIPP during disinfection,puncture,catheter insertion,and cleansing were significantly lower in the observation group than in the control group(P<0.05);the average blood oxygen saturation in the observation group was higher than that in the control group,with a lower difference compared to the control group(P<0.05);the difference and average heart rate during catheterization in the observation group were both lower than those in the control group(P<0.05);the success rates of single puncture and single catheter insertion,as well as the incidence of hypothermia post-catheterization in the observation group,were both lower than those in the control group(P<0.05),while the comparison of complication rates between the two groups showed no statistically significant difference(P>0.05).Conclusion Implementing temperature intervention during PICC placement in premature infants can effectively reduce pain,maintain stable vital signs,improve the success rates of single puncture and catheter insertion,and effectively prevent the incidence of hypothermia.

Objective : To identify autophagy-related genes involved in pulmonary infection caused by the highly drug-resistant and virulent methicillin-resistant Staphylococcus aureus strain USA300 ( USA300) ，and to explore the underlying molecular mechanisms ， thereby providing potential targets for immunotherapy． Methods: The GSE220943 dataset of a USA300-induced pulmonary infection mouse model was obtained from the GEO database． Differentially expressed genes ( DEGs ) were identified using the DESeq2 package． Autophagy-related genes ( ARGs) were retrieved from the MSigDB and Autophagy databases．Weighted gene co-expression network analysis ( WGCNA) was performed to construct gene co-expression modules．Genes overlapping among DEGs，ARGs，and WGCNA modules were identified as autophagy-related DEGs．Gene Ontology ( GO) enrichment analysis was con- ducted using the clusterProfiler R package，while Kyoto Encyclopedia of Genes and Genomes ( KEGG) pathway en- richment analysis was performed via the Metascape platform．Immune cell infiltration was analyzed using the Immu- CellAI-mouse website．A protein - protein interaction ( PPI) network was constructed using the STRING database， and hub genes were identified through topological analysis in Cytoscape． Receiver operating characteristic curve ( ROC) curves were plotted via the website https: / /www．bioinformatics．com．cn． Finally，key gene expression was validated in mouse lung tissues by real-time quantitative reverse transcription PCR ( RT-qPCR) ． Results: A total of 6 135，4 075，3 680，and 2 342 differentially expressed genes ( DEGs) were identified at 12，24，48，and 96 hours post-infection，respectively．By integrating DEGs，autophagy-related genes ( ARGs) ，and WGCNA mod- ules，19 autophagy-related DEGs were identified． GO and KEGG enrichment analyses indicated that these genes were mainly involved in CD4 + T cell activation and regulation，innate immune responses，and autophagosome mem- brane formation．Immune infiltration analysis revealed that innate immune cells such as neutrophils and dendritic cells predominated during the early phase of infection，while γδ T cells and M2 macrophages became more promi- nent in the later stages．PPI network analysis identified 12 hub autophagy-related genes，among which three upreg- ulated key genes ( Eif2ak2，Ikbke，and Nfkbiz) were further confirmed．The area under the ROC curve for all three genes was 1. 000．RT-qPCR validation demonstrated significantly elevated expression of these three genes in lung tissues at 24 hours post-infection ( all P＜0. 05) ． Conclusion Eif2ak2，Ikbke，and Nfkbiz may be involved in the pulmonary infection caused by USA300 by promoting autophagy and hold promise as potential targets for immuno- therapy．

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