OBJECTIVES: To investigate the therapeutic mechanism of nodakenin for Crohn's disease (CD)-like colitis in mice. METHODS: Using a colonic organoid model with lipopolysaccharide (LPS)- and ATP-induced pyroptosis, we investigated the effects of nodakenin on pyroptosis, intestinal barrier function and inflammatory response by detecting key pyroptosis-regulating factors and assessing changes in permeability and pro-inflammatory factors. In a mouse model of 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced CD-like colitis, the therapeutic effect of nodakenin was evaluated by measuring changes in body weight, DAI score, colonic histopathologies, inflammation score, intestinal barrier function and intestinal epithelial cell pyroptosis. The mechanism of nodakenin protection against pyroptosis of intestinal epithelial cells was explored using network pharmacology analysis and in vivo and in vitro experiments. RESULTS: In LPS- and ATP-induced colonic organoids, treatment with nodakenin significantly inhibited the expressions of NLRP3, GSDMD-N, cleaved caspase-1 and caspase-11, improved intestinal FITC-dextran (FD4, 4000) permeability, and decreased the levels of IL-1β and IL-18. In the mouse model of TNBS-induced colitis, nodakenin treatment significantly alleviated weight loss, reduced DAI score, inflammatory cell infiltration and inflammation score, and decreased serum FD4 and I-FABP levels and bacteria translocation to the mesenteric lymph nodes, spleen and liver. The mice with nodakenin treatment had also lowered expressions of NLRP3, GSDMD-N, cleaved caspase-1 and caspase-11 in the intestinal mucosa. Network pharmacology analysis suggested that the inhibitory effect of nodakenin on colitis was associated with the PI3K/Akt pathway. In both the colonic organoid model and mouse models of colitis, nodakenin effectively inhibited the activation of the PI3K/Akt pathway, and the application of IGF-1, a PI3K/Akt pathway activator, strongly attenuated the protective effect of nodakenin against intestinal epithelial cell pyroptosis and intestinal barrier dysfunction. CONCLUSIONS Nodakenin protects intestinal barrier function and alleviates CD-like colitis in mice at least partly by inhibiting PI3K/Akt signaling to reduce intestinal epithelial cell pyroptosis.
Animals ; Pyroptosis/drug effects* ; Mice ; Trinitrobenzenesulfonic Acid ; Colitis/drug therapy* ; Epithelial Cells/drug effects* ; Intestinal Mucosa/cytology* ; Disease Models, Animal ; Coumarins/pharmacology* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Crohn Disease/drug therapy*

OBJECTIVES: To investigate the therapeutic effects of cimifugin on Crohn's disease (CD)-like colitis in mice and its possible mechanism. METHODS: Thirty adult male C57BL/6 mice were randomized equally into control group, 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced CD-like colitis model group, and cimifugin treatment (daily gavage at 12.5 mg/kg) group. The therapeutic effect of cimifugin was evaluated by observing changes in body weight, disease activity index (DAI) scores, colon length, histopathological inflammation scores, and inflammatory cytokine levels in the colonic mucosa. Intestinal barrier integrity in the mice was assessed using immunofluorescence assay and Western blotting for claudin-1 and ZO-1; T-helper (Th) cell subset ratios in the mesenteric lymph nodes were analyzed with flow cytometry. Network pharmacology, KEGG enrichment analysis and molecular docking were used to predict the targets of cimifugin and analyze the key pathways and cimifugin-MAPK protein interactions, which were validated by Western blotting in the mouse models. RESULTS: In mice with TNBS-induced colitis, cimifugin treatment significantly attenuated body weight loss and colon shortening, lowered DAI and histopathological scores, decreased IFN-γ and IL-17 levels, and increased IL-4 and IL-10 levels in the colonic mucosa. Cimifugin treatment also significantly improved TNBS-induced claudin-1 dislocation and reduction of goblet cells, upregulated claudin-1 and ZO-1 expressions, reduced Th1 and Th17 cell percentages, and increased Th2 and Treg cell percentages in the colonic mucosa of the mice. KEGG analysis suggested a possible connection between the effect of cimifugin and MAPK signaling, and molecular docking showed strong binding affinity between cimifugin and MAPK core proteins. Western blotting demonstrated significantly decreased phosphorylation levels of JNK, ERK, and p38 in the colonic mucosa of cimifugin-treated mouse models. CONCLUSIONS Cimifugin alleviates TNBS-induced CD-like colitis by repairing intestinal barrier damage and restoring Th1/Th2 and Th17/Treg balance via suppressing MAPK pathway activation.
Animals ; Mice, Inbred C57BL ; Male ; Mice ; Crohn Disease/immunology* ; Colitis/immunology* ; MAP Kinase Signaling System/drug effects* ; Trinitrobenzenesulfonic Acid ; T-Lymphocytes, Helper-Inducer/drug effects* ; Intestinal Mucosa ; Disease Models, Animal

OBJECTIVES: To investigate the effect of ecliptasaponin A (ESA) for alleviating dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) in mice and the underlying mechanism. METHODS: Twenty-four male C57BL/6 mice (8-10 weeks old) were equally randomized into control group, DSS-induced IBD model group, and DSS+ESA (50 mg/kg) treatment group. Disease activity index (DAI), colon length and spleen index of the mice were measured, and intestinal pathology was examined with HE staining. The expressions of inflammatory mediators (TNF-α, IL-6, and iNOS) in the colon mucosa were detected using ELISA and RT-qPCR, and intestinal barrier integrity was assessed using AB-PAS staining and by detecting ZO-1 and claudin-1 expressions using immunofluorescence staining and Western blotting. In cultured RAW264.7 macrophages, the effects of treatment with 50 μmol/L ESA, alone or in combination with 20 μmol/L RO8191 (a JAK2/STAT3 pathway activator), on M1 polarization of the cells induced by LPS and IFN-γ stimulation and expressions of JAK2/STAT3 pathway proteins were analyzed using flow cytometry and Western blotting. RESULTS: In the mouse models of DSS-induced IBD, ESA treatment significantly alleviated body weight loss and colon shortening, reduced DAI, spleen index and histological scores, and ameliorated inflammatory cell infiltration in the colon tissue. ESA treatment also suppressed TNF‑α, IL-6 and iNOS expressions, protected the goblet cells and the integrity of the mucus and mechanical barriers, and upregulated the expressions of ZO-1 and claudin-1. ESA treatment obviously decreased CD86⁺ M1 polarization in the mesenteric lymph nodes of IBD mice and in LPS and IFN-γ-induced RAW264.7 cells, and significantly reduced p-JAK2 and p-STAT3 expressions in both the mouse models and RAW264.7 cells. Treatment with RO8191 caused reactivation of JAK2/STAT3 and strongly attenuated the inhibitory effect of ESA on CD86⁺ polarization in RAW264.7 cells. CONCLUSIONS ESA alleviates DSS-induced colitis in mice by suppressing JAK2/STAT3-mediated M1 macrophage polarization and mitigating inflammation-driven intestinal barrier damage.
Animals ; Mice ; Janus Kinase 2/metabolism* ; STAT3 Transcription Factor/metabolism* ; Mice, Inbred C57BL ; Male ; Dextran Sulfate ; Macrophages/cytology* ; Colitis/metabolism* ; Saponins/pharmacology* ; Signal Transduction/drug effects* ; RAW 264.7 Cells ; Triterpenes/pharmacology* ; Interleukin-6/metabolism*

Objective To analyze the expression level of ATP5A1 in gastric carcinoma and its influence on the prognosis of the patients and glucose metabolism in the tumor cells.Methods We retrospectively analyzed the data of 115 patients undergoing radical resection of gastric carcinoma in our hospital from February,2013 to November,2016.ATP5A1 expression in the surgical specimens were detected using immunohistochemistry,and the long-term prognosis of the patients with high(n=58)and low ATP5A1 expression(n=57)were analyzed.In gastric carcinoma MGC803 cells,the effects of lentivirus-mediated ATP5A1 knockdown or overexpression on glucose metabolism were investigated.We also observed the growth and glucose metabolism of xenografts derived from MGC803 cells with ATP5A1 knockdown or overexpression in nude mice.Results ATP5A1 was significantly overexpressed in gastric carcinoma tissues in close correlation with blood CEA and CA19-9 levels,pathological grade,T stage and N stage(P<0.05).ATP5A1 overexpression was an independent risk factor for a significantly lowered 5-year survival rate of patients with gastric carcinoma(P<0.05).ROC curve analysis demonstrated the predictive value of high ATP5A1 expression for the patients'prognosis(P<0.001).In MGC803 cells,ATP5A1 overexpression significantly up-regulated cellular glucose uptake and lactate production and increased the protein levels of HK2,PFK1,and LDHA(P<0.05),while ATP5A1 knockdown produced the opposite changes(P<0.05).In the tumor-bearing mice,overexpression of ATP5A1 increased glucose metabolism of the tumor cells and promoted tumor growth(P<0.05).Overexpression of ATP5A1 promoted the expressions of p-JNK and p-JUN in MGC803 cells(P<0.05),and the JNK inhibitor SP600125 significantly inhibited the enhancement of cellular glucose metabolism induced by ATP5A1 overexpression(P<0.05).Conclusion High ATP5A1 expression in gastric cancer is associated a poor long-term prognosis of the patients,and its effect is mediated at least partly by promoting glucose metabolism of the cells through the JNK/JUN pathway.

Objective To analyze the expression level of ATP5A1 in gastric carcinoma and its influence on the prognosis of the patients and glucose metabolism in the tumor cells.Methods We retrospectively analyzed the data of 115 patients undergoing radical resection of gastric carcinoma in our hospital from February,2013 to November,2016.ATP5A1 expression in the surgical specimens were detected using immunohistochemistry,and the long-term prognosis of the patients with high(n=58)and low ATP5A1 expression(n=57)were analyzed.In gastric carcinoma MGC803 cells,the effects of lentivirus-mediated ATP5A1 knockdown or overexpression on glucose metabolism were investigated.We also observed the growth and glucose metabolism of xenografts derived from MGC803 cells with ATP5A1 knockdown or overexpression in nude mice.Results ATP5A1 was significantly overexpressed in gastric carcinoma tissues in close correlation with blood CEA and CA19-9 levels,pathological grade,T stage and N stage(P<0.05).ATP5A1 overexpression was an independent risk factor for a significantly lowered 5-year survival rate of patients with gastric carcinoma(P<0.05).ROC curve analysis demonstrated the predictive value of high ATP5A1 expression for the patients'prognosis(P<0.001).In MGC803 cells,ATP5A1 overexpression significantly up-regulated cellular glucose uptake and lactate production and increased the protein levels of HK2,PFK1,and LDHA(P<0.05),while ATP5A1 knockdown produced the opposite changes(P<0.05).In the tumor-bearing mice,overexpression of ATP5A1 increased glucose metabolism of the tumor cells and promoted tumor growth(P<0.05).Overexpression of ATP5A1 promoted the expressions of p-JNK and p-JUN in MGC803 cells(P<0.05),and the JNK inhibitor SP600125 significantly inhibited the enhancement of cellular glucose metabolism induced by ATP5A1 overexpression(P<0.05).Conclusion High ATP5A1 expression in gastric cancer is associated a poor long-term prognosis of the patients,and its effect is mediated at least partly by promoting glucose metabolism of the cells through the JNK/JUN pathway.

Aim To explore the correlation between atherogenic index of plasma(AIP),triglyceride-glucose(TyG)index and their severity in patients with chronic coronary syndrome(CCS).Methods A total of 298 patients diagnosed with CCS by coronary angiography were retrospectively selected from Cardiology Department of the First Hospital of Lanzhou University,from May 2017 to May 2023.Clinical indexes were collected and Gensini scores were calculated based on the results of the coronary angiography,clinical data of different Gensini integral groups was compared.Linear regression was used to analyze factors that influence the elevation of Gensini scores,and receiver operator characteristic(ROC)curve was used to determine the predictive value of AIP and TyG index for the severity of coronary artery lesions in CCS.Results The research sample consisted mostly of males(77.9％)with a mean age of(61.9±8.0)years.The adjusted AIP(aAIP)and TyG index of high Gensini score group were higher than those of low Gensini score group and medium Gensini score group,and the differences were significant.Linear regression analysis revealed that total cholesterol(TC),high density lipoprotein cholesterol(HDLC),aAIP and TyG index were the factors influencing the ele-vated Gensini scores(all P＜0.05).Correlation analysis revealed that aAIP and TyG index were negatively correlated with left ventricular ejection fraction(LVEF),positively correlated with Gensini scores,and positively correlated with stent implantation and the number of stent implantation(all P＜0.05).The ROC curve results indicated that when the aAIP threshold was 1.924,the area under the ROC curve(AUC)for predicting Gensini score ≥41 points was 0.583(95％CI 0.525～0.640),with a sensitivity of 92.62％,a specificity of 25.50％,and Yoden index of 0.181.When the TyG in-dex threshold was 8.748,the AUC for predicting Gensini score ≥41 points was 0.768(95％CI 0.716～0.815),with a sensitivity of 77.18％,specificity of 67.11％,and Yoden index of 0.443.Conclusion The aAIP and TyG index were positively correlated with the severity of coronary artery disease in CCS patients.Both elevated levels can predict the severity of coronary lesions in CCS patients,but TyG index showed superior predictive ability compared with aAIP.

Objective:To analyze the monitoring results of human brucellosis prevention and control project in Gansu Province, in order to provide reference for the next revision of prevention and control plans.Methods:The data from the "China Disease Prevention and Control Information System" in Gansu Province from 2016 to 2022 and the annual data of human brucellosis cases reported by the active monitoring reports in various project counties (districts) were input into Excel 2016 software and a database was established. The epidemiological situation of brucellosis, the coverage of brucellosis prevention and control projects, the number of detected cases and health education were analyzed and compared by descriptive epidemiological method.Results:From 2016 to 2022, 70 to 82 out of 86 counties (districts) in Gansu Province were found to have human outbreaks of brucellosis, with brucellosis prevention and control projects covering 17 to 43 counties (districts). A total of 19 855 cases of human brucellosis were reported online in the province, with an average annual incidence of 11.07/100 000 and an annual incidence ranging from 6.03/100 000 to 20.89/100 000. There was a statistically significant difference in incidence among years (χ 2 = 5 264.84, P < 0.001). Through active monitoring, a total of 433 303 cases of brucellosis (tiger red plate agglutination test, RBT) were initially screened in project counties (districts) throughout the province, with 13 277 positive cases and a positive rate of 3.06% (13 277/433 303). Then the laboratory confirmed test (test-tube agglutination test, SAT) for brucellosis was carried out, with 9 967 positive cases and a positive rate of 2.30% (9 967/433 303). The infection rate of brucellosis in the population (SAT positive) was compared between different years, and the difference was statistically significant (χ 2 = 600.12, P < 0.001). In the project counties (districts), 7 481 confirmed cases of brucellosis were detected by active surveillance, and 15 794 cases were reported online, accounting for 47.37% (7 481/15 794) of the total number of cases reported online. The average awareness rate of brucellosis knowledge among medical personnel increased from 70.49% before training to 88.89% after training. The average awareness rate of brucellosis prevention and control knowledge among high-risk groups increased from 63.49% before health education to 89.05% after health education, and the behavior formation rate increased from 62.84% to 86.04%. Conclusions:From 2016 to 2022, the epidemic scope of human brucellosis in Gansu Province has been expanding year by year, and the coverage of prevention and control project is relatively small. The proportion of cases detected through active screening is relatively low. It is necessary to increase special funding investment, expand project coverage, strengthen high-risk group screening, education and behavioral intervention, and follow-up management of acute stage patient, so as to identify patients as early as possible and curb the upward trend of the epidemic.

OBJECTIVES: To investigate the effects of asperosaponin VI (AVI) on intestinal epithelial cell apoptosis and intestinal barrier function in a mouse model of Crohn's disease (CD)-like colitis and explore its mechanisms. METHODS: Male C57BL/6 mice with TNBS-induced CD-like colitis were treated with saline or AVI (daily dose 150 mg/kg) by gavage for 6 days. The changes in body weight, colon length, DAI scores, and colon pathologies of the mice were observed, and the expressions of inflammatory factors and tight injunction proteins were detected using ELISA and RT-qPCR. The effects of AVI on barrier function and apoptosis of mouse intestinal epithelial cells and TNF‑α‑treated Caco-2 cells were analyzed using immunofluorescence staining, TUNEL assay, and Western blotting. Network pharmacology, TUNEL assay, and Western blotting were performed to explore and validate the therapeutic mechanisms of AVI for CD. RESULTS: In the mouse models of CD-like colitis, AVI significantly improved body weight loss, colon shortening and DAI and tissue inflammation scores, alleviated intestinal villi and goblet cell injuries, and lowered the expressions of inflammatory factors. AVI treatment significantly reduced the loss of tight junction proteins and apoptosis in both mouse intestinal epithelial cells and TNF‑α-stimulated Caco-2 cells. KEGG enrichment pathway analysis suggested that the therapeutic effect of AVI on CD was associated with inhibition of PI3K/AKT/NF-κB pathway activation, which was confirmed by lowered expressions of p-PI3K, p-AKT, and p-p65 in AVI-treated mouse models and Caco-2 cells. In Caco-2 cells, Recilisib significantly blocked the inhibitory effect of AVI on the PI3K/AKT/NF-κB pathway and TNF-α-induced apoptosis, and AKT1 knockdown experiment confirmed the role of the PI3K/AKT pathway for mediating the activation of downstream NF-κB signaling. CONCLUSIONS AVI can improve TNBS-induced CD-like colitis in mice by reducing intestinal epithelial cell apoptosis and intestinal barrier damage via inhibiting the PI3K/AKT/NF-κB signaling pathway.
Animals ; Saponins/therapeutic use* ; Mice ; Crohn Disease/metabolism* ; Apoptosis/drug effects* ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Mice, Inbred C57BL ; Male ; Humans ; Caco-2 Cells ; Phosphatidylinositol 3-Kinases/metabolism* ; NF-kappa B/metabolism* ; Colitis/drug therapy* ; Disease Models, Animal ; Epithelial Cells/drug effects* ; Trinitrobenzenesulfonic Acid ; Intestinal Mucosa/drug effects* ; Tumor Necrosis Factor-alpha/metabolism*

Objective:To learn about the epidemic dynamics and spatial epidemic characteristics of human brucellosis in Gansu Province.Methods:Data on human brucellosis in Gansu Province reported by China Disease Control and Prevention Information System from January 2016 to December 2020 were collected and analyzed by descriptive epidemiology and spatial clustering analysis.Results:A total of 10 025 cases of human brucellosis were reported in Gansu Province from 2016 to 2020, with a statistically significant difference in incidence rate between years (χ 2 = 242.86, P = 0.001). The incidence was the lowest in 2018 (6.03/100 000), and the highest in 2020 (11.39/100 000). The reported cases were concentrated in 45 - 55 years old, accounting for 34.52% (3 461/10 025); the male to female ratio was 2.91 ∶ 1.00 (7 458/2 567); farmers were the main occupation, accounting for 82.11% (8 232/10 025). Among the 86 counties (cities, districts) in Gansu Province, Yongchang County had the highest number of reported cases in 2020 (339 cases), and Sunan Yugur Autonomous County had the highest incidence in 2020 (190.89/100 000). Spatial autocorrelation analysis showed that there was significant spatial positive correlation between the incidence rate of human brucellosis in Gansu Province from 2016 to 2020 (global Moran's I > 0, Z > 1.96, P < 0.05), showing a spatial clustering distribution. The high-high clustering areas were concentrated in Yongchang County and Sunan Yugur Autonomous County. Conclusion:In Gansu Province, the main population of human brucellosis is middle-aged male farmers, and the incidence of brucellosis is spatially clustered.

Objective:To study the epidemiological characteristics of human brucellosis in high incidence areas of brucellosis in Gansu Province, and provide a guidance for precise prevention and control of brucellosis in the areas with high incidence of the disease.Methods:Using a retrospective research method, human brucellosis monitoring data from five high incidence counties of brucellosis in Gansu Province from 2018 to 2020, including Huan County (eastern), Guazhou County (western), Zhangjiachuan County (southern), Gaotai County (northern), and Jingtai County (central), were collected from monitoring database of the Central Transfer Payment Project. The epidemic characteristics of brucellosis in high incidence areas were described and analyzed.Results:From 2018 to 2020, there were a total of 787 laboratory confirmed cases of brucellosis in high incidence areas of brucellosis in Gansu Province, with a prevalence rate of 3.64%. The incidence rates for each year were 2.63% (205/7 780), 4.60% (327/7 111), and 3.78% (255/6 746), with statistically significant differences between different years (χ 2 = 41.44, P < 0.001). The prevalence rates of brucellosis in Huan County, Guazhou County, Zhangjiachuan County, Gaotai County, and Jingtai County were 0.02% (1/4 263), 3.67% (140/3 817), 4.32% (182/4 209), 4.89% (237/4 847), and 5.04% (227/4 501), respectively. There was a statistically significant difference in comparison between different regions (χ 2 = 211.59, P < 0.001). In 2018, the incidence rate was highest in Jingtai County in the central region, and scattered to Zhangjiachuan County in the south and Gaotai County in the north in 2019 and 2020. There were statistically significant differences in the prevalence rates among different genders, ages, ethnicities, educational levels, and occupational groups (χ 2 = 111.95, 212.76, 35.32, 89.59, 97.93, P < 0.001). From 2018 to 2020, the incidence of brucellosis in the < 20 (0.78%, 6.36%, 10.00%), 40 - 50 age groups (5.15%, 7.03%, 8.00%), students and young children (0.58%, 9.20%, 13.64%), and cadres (2.86%, 4.55%, 10.53%) showed an increasing trend year by year (χ 2trend = 21.55, 7.00, 30.63, 2.01, P < 0.001). Conclusions:The brucellosis in Gansu Province is still in a high level of growth stage, and the epidemic area is gradually dispersing; the cases are gradually becoming younger, and key occupational personnel are shifting from farmers and herdsmen to social workers. It is recommended to increase the scope of monitoring, increase publicity efforts, and enhance the self-protection and control awareness of key populations.