This study aims to investigate the medication rules of patented traditional Chinese medi-cine(TCM)compound formulas and molecular mechanisms of core drugs for treating sepsis using data mining and network pharmacology approaches.In the present study,we first searched the PubMed database,Web of Science database,and the China National Knowledge Infrastructure(CNKI)since the establishment of the library to April 30,2024 for the relevant literature on the treatment of sepsis by traditional Chinese medicine.The prescriptions were then statistically ana-lyzed for drug frequency and association analysis to obtain the core drugs.Then we screened the ef-fective active ingredients of the core drugs by TCMSP and other database platforms,obtained sep-sis-related genes in GeneCards and other databases,and statistically intersected targets,and predic-ted the mechanism of action of the core TCMs by subjecting the intersected targets to PPI analy-sis,GO function and KEGG pathway enrichment analysis.Finally,the relationship between key tar-gets and herbal components was examined in reverse by molecular docking method.The results showed that 64 compound formulas were obtained,with a total of 150 Chinese medicines,which were mostly sweet in taste,cold in nature,and belonged to the spleen,stomach and intestinal me-ridians.According to the association rules,the core drugs were identified as"mirabilite-peach ker-nel-rheum officinale".There were 79 intersecting targets between the core drugs and sepsis,with core targets such as IL-1β,EGFR and SRC.MAPK,TNF,IL-17 and other signaling pathways are involved to mediate inflammatory responses,apoptosis and other biological processes to exert ther-apeutic effects on sepsis.The molecular docking results indicated that the docking activity of the key targets with the main components of the drug,and sennoside E_qt has the lowest binding ener-gy and the best docking activity with SRC.In conclusion,this study showed that the prescription of Chinese medicine for sepsis is mostly based on tonifying the spleen and clearing heat.The mecha-nism of action of the core drug"mirabilite-peach kernel-rheum officinale"in the treatment of sep-sis is multilevel and multifaceted,which provides a certain theoretical basis for the treatment of sepsis by traditional Chinese medicine.

This study was aimed at predicting and analyzing the structural and functional properties of the persistence-associated secretory protein PaaX in Mycobacterium tuberculosis(M.tb),identifying its interacting partners,and elucidating its biological roles.Bioinformatics analysis revealed that PaaX comprises 240 amino acids with a molecular mass of 26.54 kDa(C1158H1866N354O334S14).The protein lacks transmembrane domains but contains a signal peptide.Its secondary structure is dominated by α-helices(53.33%),fol-lowed by random coils(36.25%)and extended strands(10.42%),thereby forming a homotetrameric spatial configuration.Potential PaaX-interacting proteins,including Rv0406c,EchA4,EchA5,HsaE,FadE8,LpqP,and End,were predicted.These candidate genes and the paaX gene were cloned into bacterial two-hybrid vectors and co-transformed into Escherichia coli BTH101 cells.Colony PCR and sequencing confirmed the accuracy of the recombinant constructs.Bacterial two-hybrid assays demonstrated direct interac-tions of PaaX with EchA4,HsaE,FadE8,and LpqP.Moreover,gradient dilution experiments indicated that the strongest binding af-finity occurred between PaaX and EchA4.AlphaFold 3 modeling further validated these interactions,thus providing high-confidence predictions of binding interfaces.Our findings revealed that PaaX,a secreted α-helix-rich protein,engages in specific interactions with key metabolic enzymes(EchA4,HsaE,and FadE8)and a lipoprotein(LpqP),thus suggesting its potential involvement in lipid metabolism,stress adaptation,and host-pathogen interactions.This study provides novel insights into PaaX's contribution to M.tb per-sistence and pathogenicity,and highlights its value as a potential target for tuberculosis diagnostics and therapeutic development.

Objective To investigate the efficacy and mechanism of the combination of cryoablation and Yangfei establish the Lewis lung cancer mouse model,and randomly divided into model group,cryoablation group,cryoablation+cisplatin group,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups,with 5 mice in each group.Sham-operation was performed in the model group,cryoablation+cisplatin group were given intraperitoneal injection of cisplatin 3 mg/kg after cryoablation,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups were given 1.64,3.28,6.56 g/kg Yangfei Prescription by gavage after cryoablation respectively,and the model group was given equal volume of normal saline by gavage for 14 consecutive days.Tumor volume and tumor mass were measured,the morphology of tumor tissue was observed by HE staining,the expression of Ki-67 protein in tumor tissue was detected by immunohistochemistry.The differentially expressed genes in tumor tissues after cryoablation combined with Yangfei Prescription intervention were analyzed by whole transcriptome sequencing,GO and KEGG pathway enrichment analysis was performed on differentially expressed genes,and the lncRNA/circNA-mediated ceRNA network was constructed.Differentially expressed genes were verified using RT-qPCR.Results The main active components of Yangfei Prescription were terpenoids,flavonoids and aldehydes.Compared with the model group,tumor volume and tumor mass decreased(P<0.05)in cryoablation+Yangfei Prescription low-,medium-and high-dosage groups and cryoablation+cisplatin group;cryoablation combined with Yangfei Prescription could destroy the structure of tumor tissue and inhibit cell proliferation(P<0.05).A total of 1 585 differentially expressed genes(1 160 mRNA,225 lncRNA,155 miRNA and 45 circRNA)of the model group and cryoablation+Yangfei Prescription high-doseage group were obtained.GO enrichment analysis mainly involved biological processes such as immune system response and cell proliferation,cellular components such as protein complex and cell junction,molecular functions such as transcription regulator activity and molecular function regulator;KEGG pathway enrichment mainly occurred in cancer-related pathways such as PI3K-Akt signaling pathway,Th17 cell differentiation,mTOR signaling pathway,FOXO signaling pathway,etc.The lncRNA-mediated ceRNA network was composed of 97 lncRNA,73 miRNA and 417 mRNA,and the circRNA-mediated ceRNA network was composed of 26 circRNA,68 miRNA and 157 mRNA.RT-qPCR validated the expressions of 15 differentially expressed genes,which was consistent with the sequencing results.Conclusion Cryoablation combined with Yangfei Prescription can inhibit the tumor growth of Lewis lung cancer bearing mice,destroy the structure of tumor tissue,inhibit cell proliferation,the mechanism may be related to the regulation of Gm38393/miRNA-136-5p/Zfp704 axis.

Objective To investigate the value of whole lung CT radiomics combined with clinical and conventional CT features for differentiating non-tuberculous mycobacterial pulmonary disease(NTM-PD)and pulmonary tuberculosis(PTB).Methods Fifty-three NTM-PD(NTM-PD group)and 111 PTB(PTB group)patients diagnosed by mycobacteria culture were retrospectively collected.The patients were divided into training set(n=115,including 37 cases of NTM-PD and 78 cases of PTB)and test set(n=49,including 16 cases of NTM-PD and 33 cases of PTB)at the ratio of 7∶3.Patients'clinical and pulmonary CT manifestations of lesions were analyzed using univariate and multivariate logistic regression,and the independent impact factors for differentiating NTM-PD and PTB lesions were screened.The best radiomics features were extracted and screened based on whole lung CT.Based on independent impact factors,the best radiomics features and their combination,clinical-CT,radiomics and combined models were constructed with random forest,and the differential efficacy of each model was evaluated.Results Patients'age(OR=0.264),gender(OR=0.956),immunoglobulin G(OR=3.416),C reactive protein(OR=3.418)and bronchiectasis shown on CT(OR=0.285)were all independent impact factors for differentiating NTM-PD and PTB.Twelve best radiomics features were screened based on whole lung ROI.The AUC of combined model in training set and test set was 0.915 and 0.901,respectively,both higher than that of clinical model(AUC=0.832,0.801,Z=1.340,3.710,both P＜0.05)and radiomics model(AUC=0.877,0.821,Z=-2.520,-5.240,both P＜0.05).Conclusion Whole lung CT radiomics combined with clinical and conventional CT features could effectively distinguish NTM-PD and PTB.

To observe the effect of penthorum chinense pursh(PCP)on diarrhea in weaned pig-lets,and to explore its mechanism through network pharmacology and in vivo animal experiments.Animal experiment 1 A total of 160 1-day-old piglets were randomly divided into control group,low-dose prevention group(0.25％),medium-dose prevention group(0.50％)and high-dose pre-vention group(1.00％).Starting from the 14 th day,0.25％,0.50％and 1.00％PCP were added to the basal diet of the three prevention groups and weaned.PCP was stopped on the 29th day,and the diarrhea rate of piglets was recorded for 35 d.In animal experiment 2,35 4-week-old male Kunming mice were randomly divided into 5 groups(control group,LPS group,low-dose group,medium-dose group and high-dose group)for 8 d.The low-dose group,the medium-dose group and the high-dose group were intragastrically administered with 200,400 and 800 mg/kg PCP for 7 consecutive days,respectively.The control group and the LPS group were intragastrically administered with the same amount of sterile saline.On the 8th day of the experiment,except that the Control group was intraperitoneally injected with sterile normal saline,the other groups were intraperitoneally injec-ted with the same amount of LPS(15 mg/kg)to establish an intestinal injury model.HE staining was used for ileal histopathological observation,and fluorescence quantitative PCR was used to de-tect the expression levels of inflammatory factors and tight junction protein mRNA.The TCMSP database was used to screen the active components and targets of PCP,and the GeneCards database was used to obtain the targets of diarrhea.The targets of PCP and diarrhea were imported into Li-anchuan biological cloud platform,and the Venn diagram was obtained after intersection.The pro-tein-protein interaction(PPI)network was constructed by combining Cytoscape 3.7.1 and STRING database,and GO and KEGG enrichment analysis was performed by KOBAS-i platform.The results showed that compared with the control group,the diarrhea rate of weaned piglets in the low-dose prevention group(0.25％),the medium-dose prevention group(0.50％),and the high-dose prevention group(1.00％)was significantly reduced at 28-62 d(P＜0.05).According to the prediction of network pharmacology,there were 32 corresponding targets of 145 potential com-ponents of PCP,6 332 targets of diarrhea,and 118 intersection targets.The protective mechanism of PCP in the treatment of diarrhea may be related to NF-κB and PI3k-Akt signaling pathways.Further experiments confirmed that compared with the LPS group,PCP can significantly improve the pathological state of ileum in mice,the mRNA expression level of intestinal tight junction pro-tein Occludin(P＜0.05)was reversed.At the same time,PCP also significantly down-regulated the mRNA level of NF-κB.The results showed that PCP may alleviate diarrhea in piglets through multiple targets and multiple pathways.The main mechanism may be related to the inhibition of Akt-NF-κB signaling pathway.This study is conducive to providing a theoretical basis for the clini-cal application of PCP.

Objective To investigate the value of whole lung CT radiomics combined with clinical and conventional CT features for differentiating non-tuberculous mycobacterial pulmonary disease(NTM-PD)and pulmonary tuberculosis(PTB).Methods Fifty-three NTM-PD(NTM-PD group)and 111 PTB(PTB group)patients diagnosed by mycobacteria culture were retrospectively collected.The patients were divided into training set(n=115,including 37 cases of NTM-PD and 78 cases of PTB)and test set(n=49,including 16 cases of NTM-PD and 33 cases of PTB)at the ratio of 7∶3.Patients'clinical and pulmonary CT manifestations of lesions were analyzed using univariate and multivariate logistic regression,and the independent impact factors for differentiating NTM-PD and PTB lesions were screened.The best radiomics features were extracted and screened based on whole lung CT.Based on independent impact factors,the best radiomics features and their combination,clinical-CT,radiomics and combined models were constructed with random forest,and the differential efficacy of each model was evaluated.Results Patients'age(OR=0.264),gender(OR=0.956),immunoglobulin G(OR=3.416),C reactive protein(OR=3.418)and bronchiectasis shown on CT(OR=0.285)were all independent impact factors for differentiating NTM-PD and PTB.Twelve best radiomics features were screened based on whole lung ROI.The AUC of combined model in training set and test set was 0.915 and 0.901,respectively,both higher than that of clinical model(AUC=0.832,0.801,Z=1.340,3.710,both P＜0.05)and radiomics model(AUC=0.877,0.821,Z=-2.520,-5.240,both P＜0.05).Conclusion Whole lung CT radiomics combined with clinical and conventional CT features could effectively distinguish NTM-PD and PTB.

To observe the effect of penthorum chinense pursh(PCP)on diarrhea in weaned pig-lets,and to explore its mechanism through network pharmacology and in vivo animal experiments.Animal experiment 1 A total of 160 1-day-old piglets were randomly divided into control group,low-dose prevention group(0.25％),medium-dose prevention group(0.50％)and high-dose pre-vention group(1.00％).Starting from the 14 th day,0.25％,0.50％and 1.00％PCP were added to the basal diet of the three prevention groups and weaned.PCP was stopped on the 29th day,and the diarrhea rate of piglets was recorded for 35 d.In animal experiment 2,35 4-week-old male Kunming mice were randomly divided into 5 groups(control group,LPS group,low-dose group,medium-dose group and high-dose group)for 8 d.The low-dose group,the medium-dose group and the high-dose group were intragastrically administered with 200,400 and 800 mg/kg PCP for 7 consecutive days,respectively.The control group and the LPS group were intragastrically administered with the same amount of sterile saline.On the 8th day of the experiment,except that the Control group was intraperitoneally injected with sterile normal saline,the other groups were intraperitoneally injec-ted with the same amount of LPS(15 mg/kg)to establish an intestinal injury model.HE staining was used for ileal histopathological observation,and fluorescence quantitative PCR was used to de-tect the expression levels of inflammatory factors and tight junction protein mRNA.The TCMSP database was used to screen the active components and targets of PCP,and the GeneCards database was used to obtain the targets of diarrhea.The targets of PCP and diarrhea were imported into Li-anchuan biological cloud platform,and the Venn diagram was obtained after intersection.The pro-tein-protein interaction(PPI)network was constructed by combining Cytoscape 3.7.1 and STRING database,and GO and KEGG enrichment analysis was performed by KOBAS-i platform.The results showed that compared with the control group,the diarrhea rate of weaned piglets in the low-dose prevention group(0.25％),the medium-dose prevention group(0.50％),and the high-dose prevention group(1.00％)was significantly reduced at 28-62 d(P＜0.05).According to the prediction of network pharmacology,there were 32 corresponding targets of 145 potential com-ponents of PCP,6 332 targets of diarrhea,and 118 intersection targets.The protective mechanism of PCP in the treatment of diarrhea may be related to NF-κB and PI3k-Akt signaling pathways.Further experiments confirmed that compared with the LPS group,PCP can significantly improve the pathological state of ileum in mice,the mRNA expression level of intestinal tight junction pro-tein Occludin(P＜0.05)was reversed.At the same time,PCP also significantly down-regulated the mRNA level of NF-κB.The results showed that PCP may alleviate diarrhea in piglets through multiple targets and multiple pathways.The main mechanism may be related to the inhibition of Akt-NF-κB signaling pathway.This study is conducive to providing a theoretical basis for the clini-cal application of PCP.

This study was aimed at predicting and analyzing the structural and functional properties of the persistence-associated secretory protein PaaX in Mycobacterium tuberculosis(M.tb),identifying its interacting partners,and elucidating its biological roles.Bioinformatics analysis revealed that PaaX comprises 240 amino acids with a molecular mass of 26.54 kDa(C1158H1866N354O334S14).The protein lacks transmembrane domains but contains a signal peptide.Its secondary structure is dominated by α-helices(53.33%),fol-lowed by random coils(36.25%)and extended strands(10.42%),thereby forming a homotetrameric spatial configuration.Potential PaaX-interacting proteins,including Rv0406c,EchA4,EchA5,HsaE,FadE8,LpqP,and End,were predicted.These candidate genes and the paaX gene were cloned into bacterial two-hybrid vectors and co-transformed into Escherichia coli BTH101 cells.Colony PCR and sequencing confirmed the accuracy of the recombinant constructs.Bacterial two-hybrid assays demonstrated direct interac-tions of PaaX with EchA4,HsaE,FadE8,and LpqP.Moreover,gradient dilution experiments indicated that the strongest binding af-finity occurred between PaaX and EchA4.AlphaFold 3 modeling further validated these interactions,thus providing high-confidence predictions of binding interfaces.Our findings revealed that PaaX,a secreted α-helix-rich protein,engages in specific interactions with key metabolic enzymes(EchA4,HsaE,and FadE8)and a lipoprotein(LpqP),thus suggesting its potential involvement in lipid metabolism,stress adaptation,and host-pathogen interactions.This study provides novel insights into PaaX's contribution to M.tb per-sistence and pathogenicity,and highlights its value as a potential target for tuberculosis diagnostics and therapeutic development.

Objective To investigate the efficacy and mechanism of the combination of cryoablation and Yangfei establish the Lewis lung cancer mouse model,and randomly divided into model group,cryoablation group,cryoablation+cisplatin group,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups,with 5 mice in each group.Sham-operation was performed in the model group,cryoablation+cisplatin group were given intraperitoneal injection of cisplatin 3 mg/kg after cryoablation,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups were given 1.64,3.28,6.56 g/kg Yangfei Prescription by gavage after cryoablation respectively,and the model group was given equal volume of normal saline by gavage for 14 consecutive days.Tumor volume and tumor mass were measured,the morphology of tumor tissue was observed by HE staining,the expression of Ki-67 protein in tumor tissue was detected by immunohistochemistry.The differentially expressed genes in tumor tissues after cryoablation combined with Yangfei Prescription intervention were analyzed by whole transcriptome sequencing,GO and KEGG pathway enrichment analysis was performed on differentially expressed genes,and the lncRNA/circNA-mediated ceRNA network was constructed.Differentially expressed genes were verified using RT-qPCR.Results The main active components of Yangfei Prescription were terpenoids,flavonoids and aldehydes.Compared with the model group,tumor volume and tumor mass decreased(P<0.05)in cryoablation+Yangfei Prescription low-,medium-and high-dosage groups and cryoablation+cisplatin group;cryoablation combined with Yangfei Prescription could destroy the structure of tumor tissue and inhibit cell proliferation(P<0.05).A total of 1 585 differentially expressed genes(1 160 mRNA,225 lncRNA,155 miRNA and 45 circRNA)of the model group and cryoablation+Yangfei Prescription high-doseage group were obtained.GO enrichment analysis mainly involved biological processes such as immune system response and cell proliferation,cellular components such as protein complex and cell junction,molecular functions such as transcription regulator activity and molecular function regulator;KEGG pathway enrichment mainly occurred in cancer-related pathways such as PI3K-Akt signaling pathway,Th17 cell differentiation,mTOR signaling pathway,FOXO signaling pathway,etc.The lncRNA-mediated ceRNA network was composed of 97 lncRNA,73 miRNA and 417 mRNA,and the circRNA-mediated ceRNA network was composed of 26 circRNA,68 miRNA and 157 mRNA.RT-qPCR validated the expressions of 15 differentially expressed genes,which was consistent with the sequencing results.Conclusion Cryoablation combined with Yangfei Prescription can inhibit the tumor growth of Lewis lung cancer bearing mice,destroy the structure of tumor tissue,inhibit cell proliferation,the mechanism may be related to the regulation of Gm38393/miRNA-136-5p/Zfp704 axis.

This study aims to investigate the medication rules of patented traditional Chinese medi-cine(TCM)compound formulas and molecular mechanisms of core drugs for treating sepsis using data mining and network pharmacology approaches.In the present study,we first searched the PubMed database,Web of Science database,and the China National Knowledge Infrastructure(CNKI)since the establishment of the library to April 30,2024 for the relevant literature on the treatment of sepsis by traditional Chinese medicine.The prescriptions were then statistically ana-lyzed for drug frequency and association analysis to obtain the core drugs.Then we screened the ef-fective active ingredients of the core drugs by TCMSP and other database platforms,obtained sep-sis-related genes in GeneCards and other databases,and statistically intersected targets,and predic-ted the mechanism of action of the core TCMs by subjecting the intersected targets to PPI analy-sis,GO function and KEGG pathway enrichment analysis.Finally,the relationship between key tar-gets and herbal components was examined in reverse by molecular docking method.The results showed that 64 compound formulas were obtained,with a total of 150 Chinese medicines,which were mostly sweet in taste,cold in nature,and belonged to the spleen,stomach and intestinal me-ridians.According to the association rules,the core drugs were identified as"mirabilite-peach ker-nel-rheum officinale".There were 79 intersecting targets between the core drugs and sepsis,with core targets such as IL-1β,EGFR and SRC.MAPK,TNF,IL-17 and other signaling pathways are involved to mediate inflammatory responses,apoptosis and other biological processes to exert ther-apeutic effects on sepsis.The molecular docking results indicated that the docking activity of the key targets with the main components of the drug,and sennoside E_qt has the lowest binding ener-gy and the best docking activity with SRC.In conclusion,this study showed that the prescription of Chinese medicine for sepsis is mostly based on tonifying the spleen and clearing heat.The mecha-nism of action of the core drug"mirabilite-peach kernel-rheum officinale"in the treatment of sep-sis is multilevel and multifaceted,which provides a certain theoretical basis for the treatment of sepsis by traditional Chinese medicine.