AIM To explore the relieving effect of Er Miao Wan on atopic dermatitis in mice.METHODS In vivo experiment:BALB/c mice were randomly divided into normal group,model group,dexamethasone group(2 mg/kg)and high,medium and low dose groups of Er Miao Wan(4.68,2.34 and 1.17 g/kg).The mouse model of atopic dermatitis was established by repeatedly smearing DNCB solution,and the model was given orally for 21 days.The skin lesion condition on the back of mice,ear swelling degree,and the weight difference between ear lobes were observed and recorded.HE staining was used to observe the histopathological changes in the skin lesion tissues of mice.Toluidine blue(TB)staining was used to observe the infiltration of mast cells in skin lesions.The expression of macrophage marker F4/80 in skin lesions was detected by IHC.The serum levels of TSLP,IL-4,IL-5 and total IgE were detected by ELISA.In vitro experiment:RAW264.7 cells in logarithmic growth period were given 400,200 and 100 μg/mL Er Miao Wan for intervention.Cell proliferation was detected by CCK-8 method.NO level in cell supernatant was detected by Griess method.TNF-α,IL-1 β and IL-6 levels in cell supernatant were detected by ELISA method.The expressions of proteins related to the MAPK/NF-κB signaling pathway in cells was detected by Western blot.RESULTS In vivo experiment:Compared with the model group,the scores of back skin lesions,the swelling degree of right ear and the weight difference between left and right ear pieces in the high-dose group of Er Miao Wan decreased(P＜0.05,P＜0.01),the thickness of skin lesions decreased,the infiltration of mast cells and macrophages decreased(P＜0.05,P＜0.01),and the inflammatory factors TSLP,IL-4,IL-5 and total IgE levels in serum decreased(P＜0.05,P＜0.01),and the expression of F4/80 in the skin lesions decreased(P＜0.01).In vitro experiment:Compared with the model group,the levels of NO,TNF-α,IL-1 β and IL-6 in Er Miao Wan 400 and 200 μg/mL groups decreased(P＜0.05,P＜0.01),and the phosphorylation levels of P38,JNK and P65 proteins decreased(P＜0.05,P＜0.01).CONCLUSION Er Miao Wan can alleviate skin lesion inflammation in DNCB-induced atopic dermatitis mice,and its mechanism may be related to inhibiting the activation of MAPK/NF-κB signaling pathway of macrophage,reducing macrophage infiltration and reducing Th2 cytokines.

Objective:To investigate the effect of securinine(SEC)on apoptosis of the human colon cancer cell line SW620,and to elucidate its possible mechanism.Methods:The nude mice with subcutaneously transplanted tumor were divided into control group(n=6),oxaliplatin(OXA)group(n=7),and SEC group(n=7).The volume and mass of subcutaneous tumors in the nude mice were measured in various groups,and the tumor inhibitory rates in various groups were calculated.The SW620 cells were treated with different doses(5-120 μmol·L-1)of SEC for 12,24,48,and 72 h,respectively.Cell counting kit-8(CCK-8)assay was used to assess the survival rates of cells in various groups,and the optimal doses of SEC were confirmed.The SW620 cells were divided into control group,20 μmol·L-1 SEC group,40 μmol·L-1SEC group,and 40 μmol·L-1OXA group.TUNEL staining method and flow cytometry were used to detect the apoptotic rates of cells in various groups.JC-1 staining was used to detect the mitochondrial membrane potentials of cells in various groups,and 2',7'-dichlorodi-hydrofluorescin diacetate(DCFH-DA)fluorescence staining and flow cytometry were used to measure the reactive oxygen species(ROS)levels in the cells in various groups.Western blotting method was used to detect the expression levels of cytochrome C,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),c-Jun N-terminal kinase(JNK),phosphorylated JNK(p-JNK),mitogen-activated protein kinase p38,phosphorylated p38(p-p38),extracellular signal-regulated kinase(ERK)and phosphorylated ERK(p-ERK)proteins in the cells in various groups.Results:Compared with control group,the volume and mass of subcutaneously transplanted tumors in the nude mice in SEC group were significantly decreased(P＜0.05 or P＜0.01 or P＜0.001);the inhibitory rates of tumor in SEC group and OXA group were 20.42％and 6.50％.The CCK-8 assay results showed that compared with 0 μmol·L-1 SEC,when the SEC dose exceeded 20 μmol·L-1,the survival rates of SW620 cells were significantly decreased(P＜0.001).The optimal condition for subsequent experiments was set as doses of 20 μmol·L-1SEC and 40 μmol·L-1SEC,and duration of 24 h.The TUNEL results showed that compared with control group,the apoptotic rates of cells in 20 and 40 μmol·L-1 SEC groups were significantly increased(P＜0.05 or P＜0.001).The results of flow cytometry showed that compared with control group,the apoptotic rate in 40 μmol·L-1SEC group was significantly increased(P＜0.001).The JC-1 staining results showed that compared with control group,the mitochondrial membrane potentials of cells in 20 and 40 μmol·L-1 SEC groups were significantly decreased(P＜0.001).Compared with control group,the levels of ROS detected by DCFH-DA fluorescence staining in the cells of 20 and 40 μmol·L-1 SEC groups and 40 μmol·L-1 OXA group were significantly increased(P＜0.001),while the level of ROS detected by flow cytometry in 40 μmol·L-1SEC group was significantly increased(P＜0.05).Compared with control group,the expression levels of Bcl-2 protein in the cells in 20 and 40 μmol·L-1 SEC groups and 40 μmol·L-1 OXA group were decreased(P＜0.01),while the expression levels of cytochrome C and Bax proteins were increased(P＜0.001).Compared with control group,the ratios of p-JNK/JNK,p-p38/p38 and p-ERK/ERK in 20 and 40 μmol·L-1 SEC groups were significantly increased(P＜0.05 or P＜0.01 or P＜0.001).Conclusion:SEC can inhibit the proliferation of SW620 cells,increase the cellular ROS levels,reduce the mitochondrial membrane potential,and induce the mitochondrial apoptosis;its mechanism may be related to the regulation of the mitogen-activated protein kinase(MAPK)signaling pathway.

Objective:To discuss the regulatory role of complement alternative pathway in mouse colorectal cancer(CRC)liver metastasis model based on bioinformatics methods,and to clarify its mechanism through experimental verification.Methods:Using"CRC liver metastasis"as the keyword,the GSE81558 dataset was retrieved from Gene Expression Omnibus(GEO)database,including normal colon tissue samples,CRC tissue samples and CRC liver metastasis tissue samples.Bioinformatics methods were used to analyze and screen differentially expressed genes(DEGs).Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using R and Cytoscape software,and the results were visualized.Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database was used to evaluate protein-protein interactions(PPIs)of DEGs and construct PPI network.Twelve C57BL/6 mice were injected with SL4 tumor cells into spleen,and the liver tissues were collected at 0,7 and 14 d.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of complement pathway-related genes in liver metastatic foci.The CRC liver metastasis mouse model was used to verify the complement signaling pathway.The mice were divided into control group,factor B knockout group(FB-/-)and C4 factor knockout group(C4-/-),and there were 6 mice in each group.The liver weights of the mice were measured;HE staining was used to detect the percentage of metastatic area in liver tissue in control group and FB-/-group;immunohistochemistry was used to detect macrophage infiltration in liver tissue in control group and FB-/-group,and the percentage of macrophage infiltration was calculated.Results:The distances between normal colon tissue samples and CRC tissue samples,as well as between CRC tissue samples and CRC liver metastasis tissue samples were far,indicating significant differences between samples,allowing subsequent analysis of DEGs.A total of 1 908 DEGs were screened in the dataset comparing normal colon tissue samples and CRC tissue samples,including 771 up-regulated DEGs and 1 137 down-regulated DEGs.Twenty-three up-regulated DEGs and 100 down-regulated DEGs were identified in the dataset comparing CRC and CRC liver metastasis.The GO functional enrichment analysis results showed that compared with normal colon tissue samples,DEGs in CRC samples were mainly enriched in biological processes(BP)related to cell cycle and mitosis,including mitotic cell cycle process,cell division,response to hormone,mitotic nuclear division and response to lipid.Compared with CRC samples,the DEGs in CRC liver metastasis samples were mainly enriched in coagulation-related BP,including platelet degranulation,blood coagulation regulation,acute-phase response,hemostasis regulation and coagulation regulation.The KEGG pathway enrichment analysis results showed that compared with normal colon tissue samples,the DEGs in CRC tissue samples were mainly enriched in cell cycle and p53 signaling pathways.Compared with CRC tissue samples,the DEGs in CRC liver metastasis tissue samples were mainly enriched in complement,coagulation cascade and metabolism-related signaling pathways.The Hub genes identified in PPI network were related to blood proteins.The RT-qPCR results showed that compared with 0 d group,the mRNA expression level of complement related genes complement 1q(C1q)in liver metastatic foci tissue sampres in 7 d group was significantly decreased(P<0.05),the mRNA expression levels of complement 3(C3),complement 5(C5),FB,and factor D(FD)were significantly increased(P<0.05 or P<0.01),the mRNA expression levels of complement pathway-related genes C1q,complement 2(C2),C3,complement fragment 3a receptor(C3aR),C5,complement fragment 5a receptor(C5aR),decay-accelerating factor(DAF),FB and FD in liver metastatic foci tissue sampres in 14 d group were significantly increased(P<0.05 or P<0.01).Compared with control group,the liver weight of the mice in FB-/-group was significantly decreased(P<0.01),while there was no significant difference was observed in C4-/-group(P>0.05).The HE staining results showed that compared with control group,the liver metastatic foci in FB-/-mice were significantly decreased,and the percentage of metastatic area was decreased(P<0.01).The immunohistochemistry results showed that compared with control group,the macrophage infiltration in liver metastatic foci of the mice in FB-/-group was reduced,and the percentage of macrophage infiltration was decreased(P<0.01).Conclusion:Complement cascade is associated with CRC liver metastasis,and the alternative complement pathway regulates CRC liver metastasis,suggesting this pathway may serve as a potential therapeutic target for CRC liver metastasis.

Objective To investigate the interaction between a novel antimicrobial compound,HL-J6,and penicillin-binding protein 1(PBP1)of Staphylococcus aureus.Methods With MRSA252 genomic DNA as the template and PBP1F and PBP1R as primers,the expression plasmid pET30a-pbp1-39-608 was constructed by amplifying the target gene fragment followed by cloning into the Nde I/Xho I restriction sites of the pET30a vector.Then the obtained plasmids were transformed into Escherichia coli for the expression of PBP1-39-608 protein,and the product was purified by affinity chromatography.The inhibitory effect of HL-J6 on the transpeptidase activity of PBP1-39-608 was measured using peptidoglycan side chain backbone peptide,with thiol ester analog S2d as the substrate.The affinity between HL-J6 and PBP1-39-608 was detected using microscale thermophoresis(MST),and the binding interaction was confirmed by cellular thermal shift assay(CETSA).Molecular docking and dynamics simulation were performed using AutoDock Vina and Desmond software,respectively,to elucidate the binding mode of HL-J6 with the PBP1-39-608 protein and the key amino acid residues involved.Results The recombinant plasmid pET30a-pbp1-39-608 was successfully constructed,and PBP1-39-608 protein was produced after induction and purified,yielding a protein with an approximate molecular mass of 65×103.HL-J6 inhibited the transpeptidase activity of PBP1-39-608 in a time-dependent manner(P<0.001).The dissociation constant Kd of the binding between HL-J6 and PBP1-39-608 was 64.92 μmol/L.Molecular docking results showed that HL-J6 bound to the active pocket of PBP1-39-608 by interacting with key residues such as ILE-348,ASN-370,THR-516 and PHE-423,with a binding score of-8.38 kcal/mol(<-5.00 kcal/mol).Dynamics simulation results indicated that the complex became stable after 50 ns.Conclusion HL-J6 effectively inhibits the transpeptidase activity of Staphylococcus aureus PBP1,and shows stable interaction with the protein.

Emodin is a hydroxyanthraquinone compound that is widely distributed and has multiple pharmacological activities, including anti-diarrheal, anti-inflammatory, and liver-protective effects. Research indicates that emodin may be one of the main components responsible for inducing hepatotoxicity. However, studies on the mechanisms of liver injury are relatively limited, particularly those related to bile acids(BAs) metabolism. This study aims to systematically investigate the effects of different dosages of emodin on BAs metabolism, providing a basis for the safe clinical use of traditional Chinese medicine(TCM)containing emodin. First, this study evaluated the safety of repeated administration of different dosages of emodin over a 5-week period, with a particular focus on its impact on the liver. Next, the composition and content of BAs in serum and liver were analyzed. Subsequently, qRT-PCR was used to detect the mRNA expression of nuclear receptors and transporters related to BAs metabolism. The results showed that 1 g·kg~(-1) emodin induced hepatic damage, with bile duct hyperplasia as the primary pathological manifestation. It significantly increased the levels of various BAs in the serum and primary BAs(including taurine-conjugated and free BAs) in the liver. Additionally, it downregulated the mRNA expression of farnesoid X receptor(FXR), retinoid X receptor(RXR), and sodium taurocholate cotransporting polypeptide(NTCP), and upregulated the mRNA expression of cholesterol 7α-hydroxylase(CYP7A1) in the liver. Although 0.01 g·kg~(-1) and 0.03 g·kg~(-1) emodin did not induce obvious liver injury, they significantly increased the level of taurine-conjugated BAs in the liver, suggesting a potential interference with BAs homeostasis. In conclusion, 1 g·kg~(-1) emodin may promote the production of primary BAs in the liver by affecting the FXR-RXR-CYP7A1 pathway, inhibit NTCP expression, and reduce BA reabsorption in the liver, resulting in BA accumulation in the peripheral blood. This disruption of BA homeostasis leads to liver injury. Even doses of emodin close to the clinical dose can also have a certain effect on the homeostasis of BAs. Therefore, when using traditional Chinese medicine or formulas containing emodin in clinical practice, it is necessary to regularly monitor liver function indicators and closely monitor the risk of drug-induced liver injury.
Emodin/administration & dosage* ; Bile Acids and Salts/metabolism* ; Animals ; Male ; Liver/injuries* ; Chemical and Drug Induced Liver Injury/genetics* ; Drugs, Chinese Herbal/adverse effects* ; Humans ; Rats, Sprague-Dawley ; Mice ; Rats

OBJECTIVES: To study the effect of prophylactic phenytoin (PHT) or levetiracetam (LEV) on busulfan (BU) blood concentration in children undergoing hematopoietic stem cell transplantation. METHODS: Pediatric patients conditioned with BU plus cyclophosphamide and fludarabine at the First People's Hospital of Chenzhou from September 2023 to February 2025 were retrospectively included. Patients were grouped by prophylactic antiepileptic regimen into PHT (n=24) and LEV (n=26). BU blood concentrations at the end of infusion (0 hour) and at 1, 2, and 4 hours post-infusion were compared between groups. RESULTS: At 0 hour post-infusion, BU blood concentrations did not differ significantly between groups (P>0.05). At 1, 2, and 4 hours post-infusion, BU blood concentrations were higher in the LEV group than in the PHT group (P<0.05). The area under the concentration-time curve from 0 to ∞ (AUC_0-∞) was greater in the LEV group (P<0.001), and the attainment rate of AUC_0-∞ was higher in the LEV group than in the PHT group (73% vs 21%, P<0.001). No significant differences were observed between groups in time to hematopoietic engraftment or in the incidence of BU-related adverse drug reactions (P>0.05). CONCLUSIONS Compared with PHT, LEV prophylaxis is associated with higher BU blood concentration and a higher AUC_0-∞ attainment rate. There is no observed difference in BU efficacy or safety between PHT and LEV.
Humans ; Levetiracetam/therapeutic use* ; Busulfan/pharmacokinetics* ; Hematopoietic Stem Cell Transplantation ; Male ; Female ; Child ; Child, Preschool ; Phenytoin/pharmacology* ; Infant ; Retrospective Studies ; Anticonvulsants/pharmacology* ; Adolescent

Objective To investigate the predictive value of serum interleukin-6(IL-6)and procalcitonin(PCT)levels on the prognosis of patients with acute respiratory distress syndrome(ARDS).Methods A total of 99 ARDS patients admitted to Lianyungang Hospital affiliated to Xuzhou Medical University from January 2021 to December 2023 were included in the study.According to the Berlin definition,the patients were divided into mild group(n=22),moderate group(n=42)and severe group(n=35)groups.Based on their 28-day outcomes,they were also divided into survival group(n=65)and death group(n=34).The levels of IL-6 and PCT in each group were compared.The predictive value of IL-6 and PCT for the prognosis of ARDS patients was analyzed using receiver operat-ing characteristic(ROC)curve and the area under the curve(AUC).The Kaplan-Meier method was used to plot survival curves for ARDS patients with different levels of IL-6 and PCT.Results Among the three groups with varying severity of ARDS,the severe group had the highest IL-6 and PCT levels,followed by the moderate group,and the mild group had the lowest levels.The differences among the groups were statistically significant(P＜0.05).The levels of IL-6 and PCT in the death group were significantly higher than those in the survival group,and the difference was statistically significant(P＜0.05).The AUC of IL-6 and PCT in predicting the prognosis of ARDS patients were 0.793 and 0.776,with the sensitivity of 79.4％and 94.1％,and the specificity of 70.8％and 61.5％,respective-ly.Kaplan-Meier survival curve analysis showed that when IL-6 was ≥110.700pg/ml and PCT was ≥ 1.700ng/ml,the 28-day sur-vival rate of ARDS patients was significantly lower than that of patients with IL-6＜110.700pg/ml and PCT＜1.700ng/ml(P＜0.001).Conclusion PCT and IL-6 levels have good value in predicting the short-term prognosis of ARDS patients,and can be used as effective indicators for prognosis evaluation in ARDS patients.

Objective To explore the safety and efficacy of percutaneous sacroiliac screw combined with retrograde pubic ramus screws in the treatment of unstable pelvic fractures.Methods A retrospective analysis was made on clinical data of 32 cases of unstable pelvic fractures treated with percutaneous sacroiliac screws combined with retrograde pubic ramus screws from August 2021 to November 2023.The channel screws were inserted under the guidance of C-arm fluoroscopy.Results A total of 75 channel screws were inserted,including 36 sacroiliac screws and 39 retrograde pubic ramus screws.Each sacroiliac screw underwent fluoroscopy for(32.2±4.6)times,and each pubic ramus screw for(40.3±11.7)times.The operation time was(154.2±43.8)min,and the intraoperative blood loss was(30.5±8.7)ml.There were no iatrogenic vascular or nerve injuries.One case of pubic ramus screw infection occurred after surgery,and a debridement was performed to remove internal fixation.All the fractures had bone union,and the healing time was(13.2±3.7)weeks.The quality of fracture reduction(Matta criteria)was excellent in 27 cases,good in 3 cases,and fair in 2 cases,with an excellent and good rate of 93.8％(30/32).CT images showed the penetration of 73 screws at level 0,1 screw at level 1,and 1 at level 2.All the 32 cases were followed up for 6-42 months(mean,16.3±6.1 months).According to the Majeed functional scoring criteria,28 cases were excellent,3 cases were good,and 1 case was fair,with an excellent and good rate of 96.9％(31/32).Conclusion Percutaneous sacroiliac screws combined with retrograde pubic ramus screws in the treatment of unstable pelvic fractures is easy to operate and safe,and has satisfactory therapeutic effects.

Objective To explore the safety and efficacy of percutaneous sacroiliac screw combined with retrograde pubic ramus screws in the treatment of unstable pelvic fractures.Methods A retrospective analysis was made on clinical data of 32 cases of unstable pelvic fractures treated with percutaneous sacroiliac screws combined with retrograde pubic ramus screws from August 2021 to November 2023.The channel screws were inserted under the guidance of C-arm fluoroscopy.Results A total of 75 channel screws were inserted,including 36 sacroiliac screws and 39 retrograde pubic ramus screws.Each sacroiliac screw underwent fluoroscopy for(32.2±4.6)times,and each pubic ramus screw for(40.3±11.7)times.The operation time was(154.2±43.8)min,and the intraoperative blood loss was(30.5±8.7)ml.There were no iatrogenic vascular or nerve injuries.One case of pubic ramus screw infection occurred after surgery,and a debridement was performed to remove internal fixation.All the fractures had bone union,and the healing time was(13.2±3.7)weeks.The quality of fracture reduction(Matta criteria)was excellent in 27 cases,good in 3 cases,and fair in 2 cases,with an excellent and good rate of 93.8％(30/32).CT images showed the penetration of 73 screws at level 0,1 screw at level 1,and 1 at level 2.All the 32 cases were followed up for 6-42 months(mean,16.3±6.1 months).According to the Majeed functional scoring criteria,28 cases were excellent,3 cases were good,and 1 case was fair,with an excellent and good rate of 96.9％(31/32).Conclusion Percutaneous sacroiliac screws combined with retrograde pubic ramus screws in the treatment of unstable pelvic fractures is easy to operate and safe,and has satisfactory therapeutic effects.

Objective To investigate the predictive value of serum interleukin-6(IL-6)and procalcitonin(PCT)levels on the prognosis of patients with acute respiratory distress syndrome(ARDS).Methods A total of 99 ARDS patients admitted to Lianyungang Hospital affiliated to Xuzhou Medical University from January 2021 to December 2023 were included in the study.According to the Berlin definition,the patients were divided into mild group(n=22),moderate group(n=42)and severe group(n=35)groups.Based on their 28-day outcomes,they were also divided into survival group(n=65)and death group(n=34).The levels of IL-6 and PCT in each group were compared.The predictive value of IL-6 and PCT for the prognosis of ARDS patients was analyzed using receiver operat-ing characteristic(ROC)curve and the area under the curve(AUC).The Kaplan-Meier method was used to plot survival curves for ARDS patients with different levels of IL-6 and PCT.Results Among the three groups with varying severity of ARDS,the severe group had the highest IL-6 and PCT levels,followed by the moderate group,and the mild group had the lowest levels.The differences among the groups were statistically significant(P＜0.05).The levels of IL-6 and PCT in the death group were significantly higher than those in the survival group,and the difference was statistically significant(P＜0.05).The AUC of IL-6 and PCT in predicting the prognosis of ARDS patients were 0.793 and 0.776,with the sensitivity of 79.4％and 94.1％,and the specificity of 70.8％and 61.5％,respective-ly.Kaplan-Meier survival curve analysis showed that when IL-6 was ≥110.700pg/ml and PCT was ≥ 1.700ng/ml,the 28-day sur-vival rate of ARDS patients was significantly lower than that of patients with IL-6＜110.700pg/ml and PCT＜1.700ng/ml(P＜0.001).Conclusion PCT and IL-6 levels have good value in predicting the short-term prognosis of ARDS patients,and can be used as effective indicators for prognosis evaluation in ARDS patients.