1.Construction of a nomogram model for clinical cure of chronic hepatitis B with a low level of hepatitis B surface antigen treated with pegylated interferon α-2b
Yingyuan ZHANG ; Huan MU ; Lixian CHANG ; Danqing XU ; Yuanzhen WANG ; Chunyun LIU ; Weikun LI ; Huangchenghao ZHANG ; Chunyan MOU ; Li LIU
Journal of Clinical Hepatology 2026;42(5):1038-1047
ObjectiveTo investigate the predictive factors for HBsAg clearance in chronic hepatitis B (CHB) patients with a low level of hepatitis B surface antigen (HBsAg) treated with pegylated interferon α-2b (PEG-IFN-α-2b), to establish a combined predictive model and a nomogram based on multiple factors, and to provide a reference for formulating individualized treatment regimens and predicting treatment outcome in clinical practice. MethodsA retrospective analysis was performed for 167 CHB patients with HBsAg <1 500 IU/mL who attended The Third People’s Hospital of Kunming from January 2022 to January 2024 and were treated with PEG-IFN-α-2b. According to whether clinical cure was achieved, the patients were divided into HBsAg clearance group and HBsAg non-clearance group. Related data were collected, including general information and serological/biochemical/virological indicators at different time points during treatment. The independent samples t-test was used for comparison of normally distributed continuous data, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data; the chi-square test was used for comparison of categorical data. The multivariate logistic regression analysis was used to identify independent influencing factors. The receiver operating characteristic (ROC) curve was used to assess the value of indicators used alone or in combination in predicting clinical cure, and calibration curves were plotted to assess the risk prediction model. ResultsThe univariate analysis showed that there were significant differences between the two groups in age (t=-6.839, P<0.05), history of nucleos(t)ide analogue treatment for over 1 year (χ2=59.339, P<0.05), genotype (χ2=4.610, P<0.05), nonalcoholic fatty liver disease (χ2=5.319, P<0.05), hepatitis B virus DNA status before treatment (χ2=60.861, P <0.05), compensated liver cirrhosis (χ2=10.960, P<0.05), HBeAg status before treatment (χ2=19.060, P<0.05), a history of interferon treatment (χ2=8.162, P<0.05), presence of interferon antibodies after treatment (χ2=12.858, P<0.05), HBsAg level before treatment (Z=-7.412, P<0.05), alanine aminotransaminase (ALT) level at baseline (Z=-6.117, P<0.05), ALT level at 12 weeks of treatment (Z=-7.171, P<0.05), platelet count (PLT) at 24 weeks of treatment (Z=-3.622, P<0.05), and thyroid stimulating hormone (TSH) level at 24 weeks of treatment (Z=-2.830, P<0.05). The multivariate logistic regression analysis showed that age (odds ratio [OR]=1.230, P=0.007), history of nucleos(t)ide analogue treatment for over 1 year (OR=0.008, P=0.011), HBeAg status before treatment (OR=0.003, P=0.012), HBsAg level before treatment (OR=1.005, P=0.014), ALT level at baseline (OR=0.949, P=0.014), ALT level at 12 weeks of treatment (OR=0.969, P=0.016), PLT at 24 weeks of treatment (OR=0.969, P=0.022), and TSH level at 24 weeks of treatment (OR=3.608, P=0.045) were independent influencing factors for HBsAg clearance at 48 weeks of treatment in CHB patients with HBsAg <1 500 IU/mL. The Hosmer-Lemeshow goodness-of-fit test yielded χ2=1.398, P=0.994, indicating that the model had good fitting. The Bootstrap method was used to perform internal validation of the nomogram model, and there was a good degree of fitting between the calibration curve and the ideal curve, with a mean absolute error of 0.029. The ROC curve analysis showed that the combination of predictive factors had an area under the ROC curve of 0.982 (95% confidence interval: 0.961 — 0.999), with a sensitivity of 94.10% and a specificity of 93.10%, suggesting that the nomogram model had a good discriminatory ability. For the CHB patients with HBsAg <1 500 IU/mL and different features, further analysis of HBsAg clearance rate at 48 weeks of treatment showed an HBsAg clearance rate of 69.60% for those with HBsAg ≤67.65 IU/mL before treatment, 58.30% for those with a baseline ALT level of ≥62.50 U/L, 68.30% for those with an ALT level of ≥92.50 U/L at 12 weeks of treatment, 42.40% for those with PLT ≥104×109/L at 24 weeks of treatment, and 48.30% for those with a TSH level of ≤1.38 μIU/mL at 24 weeks of treatment, with significant differences between the two groups (all P<0.001). ConclusionAge, history of nucleos(t)ide analogue treatment for over 1 year, HBeAg status before treatment, HBsAg level before treatment, baseline ALT level, ALT level at 12 weeks of treatment, PLT level at 24 weeks, and TSH level at 24 weeks of treatment are independent predictive factors. The combined prediction nomogram model constructed in this study has a relatively high value in predicting clinical cure at 48 weeks of PEG-IFN-α-2b treatment in CHB patients with HBsAg<1 500 IU/mL, thereby providing a reference for selecting suitable treatment population and predicting clinical cure.
2.Distribution and drug resistance of pathogens isolated from different age groups of children with urinary tract infections in a health care hospital of Guangxi Province
Minxue LIU ; Liying HUANG ; Jiahui LIANG ; Huan ZHANG ; Chunyun FU
Chinese Journal of Nosocomiology 2025;35(12):1846-1851
OBJECTIVE To analyze the situation of urinary tract infections(UTIs)in children of different age groups in a health care hospital of Guangxi,and to analyze the detected pathogens and drug resistance rate.METHOD Data on urinary tract infections in children between 2017 and 2023 in Guangxi provincial maternal and child healthcare hospitals were retrospectively analyzed,and children were classified according to age:neonates(≤28 days),infants(>28 days and ≤1 year),preschoolers(>1 year and<6 year)and 6-14 years old.The urina-ry tract infections,pathogen identification and drug resistance rates of major pathogens in children of different age groups were analyzed.RESULTS The pathogens of pediatric UTIs in each group were dominated by gram-negative bacilli(44.16%-67.36%),with the highest percentage of Escherichia coli(21.81%—38.60%),gram-posi-tive infections were dominated by Enterococcus faecium(5.96%—21.40%)and Enterococcus faecalis(4.66%—13.68%),and fungi were dominated by Candida albicans(8.03%-12.75%).Admission to intensive care unit was higher in the neonates group(37.38%,P<0.001).Urine culture positivity rate was elevated in the 6-14 years age group(31.93%,P<0.001),with girls being more common(59.47%,P<0.001).The rate of E.coli resistance to cephalosporins was relative high in urine culture isolates from the infant group(28%—54%).In ad-dition,the resistance rate of Enterococcus faecium from urine to ampicillin was higher in the infant group than that in the preschool children and 6-14 years old groups(P=0.002).CONCLUSIONS The gram-negative bacteri-a were dominant among the pathogens isolated from the children with urinary tract infections and showed certain drug resistance to the commonly used antibiotics.Urinary tract infections are more difficult to diagnose and treat in younger children,and pediatricians should pay more attention to them.
3.Distribution and drug resistance of pathogens isolated from different age groups of children with urinary tract infections in a health care hospital of Guangxi Province
Minxue LIU ; Liying HUANG ; Jiahui LIANG ; Huan ZHANG ; Chunyun FU
Chinese Journal of Nosocomiology 2025;35(12):1846-1851
OBJECTIVE To analyze the situation of urinary tract infections(UTIs)in children of different age groups in a health care hospital of Guangxi,and to analyze the detected pathogens and drug resistance rate.METHOD Data on urinary tract infections in children between 2017 and 2023 in Guangxi provincial maternal and child healthcare hospitals were retrospectively analyzed,and children were classified according to age:neonates(≤28 days),infants(>28 days and ≤1 year),preschoolers(>1 year and<6 year)and 6-14 years old.The urina-ry tract infections,pathogen identification and drug resistance rates of major pathogens in children of different age groups were analyzed.RESULTS The pathogens of pediatric UTIs in each group were dominated by gram-negative bacilli(44.16%-67.36%),with the highest percentage of Escherichia coli(21.81%—38.60%),gram-posi-tive infections were dominated by Enterococcus faecium(5.96%—21.40%)and Enterococcus faecalis(4.66%—13.68%),and fungi were dominated by Candida albicans(8.03%-12.75%).Admission to intensive care unit was higher in the neonates group(37.38%,P<0.001).Urine culture positivity rate was elevated in the 6-14 years age group(31.93%,P<0.001),with girls being more common(59.47%,P<0.001).The rate of E.coli resistance to cephalosporins was relative high in urine culture isolates from the infant group(28%—54%).In ad-dition,the resistance rate of Enterococcus faecium from urine to ampicillin was higher in the infant group than that in the preschool children and 6-14 years old groups(P=0.002).CONCLUSIONS The gram-negative bacteri-a were dominant among the pathogens isolated from the children with urinary tract infections and showed certain drug resistance to the commonly used antibiotics.Urinary tract infections are more difficult to diagnose and treat in younger children,and pediatricians should pay more attention to them.
4.Analysis of influencing factors and construction of predictive model for HBsAg clearance in patients with HBeAg-negative chronic hepatitis B treated with PEG-IFN-α-2b
Yingyuan ZHANG ; Danqing XU ; Huan MU ; Yuanqiang HE ; Yuanzhen WANG ; Chunyun LIU ; Weikun LI ; Chunyan MOU ; Li LIU
Journal of Clinical Hepatology 2025;41(8):1525-1532
Objective To investigate the predictive factors for the occurrence of HBsAg clearance in patients with HBeAg-negative chronic hepatitis B(CHB)receiving peginterferon alfa-2b(PEG-IFN-α-2b)treatment,analyze the effects of various indicators on the HBsAg clearance rate under different characteristics,and construct and evaluate a combined predictive model.Methods We included 125 patients with HBeAg-negative CHB at Kunming Third People's Hospital from May 2021 to May 2023.After treatment with PEG-IFN-α-2b combined with nucleoside analogues for a course of 48 weeks,they were divided into HBsAg clearance group and HBsAg non-clearance group.Their general information and serological,biochemical,and virological indicators at different time points during treatment were recorded.Continuous data in normal distribution were compared using the t test.Continuous data in non-normal distribution were compared using the Mann-Whitney U test,and comparisons across different time points were performed using the multiple paired-sample Friedman test.Categorical data were compared using the χ2 test.A Logistic regression analysis was used to select variables to establish a combined multi-parameter predictive model.Receiver operating characteristic(ROC)curves were generated to evaluate the diagnostic value of individual indicators and the combined predictive model for HBsAg clearance.Results Before treatment,there were significant differences in baseline HBsAg level(Z=-3.997,P<0.05)and treatment history(χ2=8.221,P<0.05)between the two groups.During treatment,gradually decreasing trends were observed in white blood cell count(χ2=104.944),neutrophil count(χ2=132.036),platelet count(χ2=162.881),and thyroid-stimulating hormone level(TSH,χ2=83.304,all P<0.05),while alanine aminotransferase(ALT,χ2=157.618)and alpha fetoprotein(χ2=159.472)showed gradually increasing trends(both P<0.05).At 48 weeks of treatment,treatment history(odds ratio[OR]=0.232,95%confidence interval[CI]:0.071-0.753),baseline HBsAg level(OR=13.423,95%CI:3.276-54.997),the extent of decrease in HBsAg from baseline after 12 weeks of treatment(OR=0.143,95%CI:0.040-0.515),the maximum ALT level during treatment(OR=0.986,95%CI:0.980-0.993),and the minimum TSH level during treatment(OR=3.281,95%CI:1.413-7.619)were independent factors affecting HBsAg clearance(all P<0.05).A combined predictive model for HBsAg clearance was built:Y=-1.603-1.462×treatment history+2.597×baseline HBsAg value-1.944×the extent of HBsAg reduction from baseline after 12 weeks of treatment-0.014×the maximum ALT value during treatment+1.188×the minimum TSH value during treatment.The diagnostic value of the individual indicators for HBsAg clearance from high to low was as following:the maximum ALT value during treatment(AUC=0.824),baseline HBsAg value(AUC=0.727),the minimum TSH value during treatment(AUC=0.707),the extent of HBsAg reduction from baseline after 12 weeks of treatment(AUC=0.641),and treatment history(AUC=0.636).The combined model showed better predictive performance than the individual indicators,with the AUC being 0.921(all P<0.05).Conclusion The combined model,constructed with baseline HBsAg value,the extent of HBsAg reduction from baseline after 12 weeks of treatment,the maximum ALT value during treatment,and the minimum TSH value during treatment,has high predictive value for the occurrence of HBsAg clearance in patients with HBeAg-negative CHB after 48 weeks of treatment with PEG-IFN-α-2b,which can provide a reference for identifying suitable patients for treatment and predicting clinical outcome.
5.Analysis of influencing factors and construction of predictive model for HBsAg clearance in patients with HBeAg-negative chronic hepatitis B treated with PEG-IFN-α-2b
Yingyuan ZHANG ; Danqing XU ; Huan MU ; Yuanqiang HE ; Yuanzhen WANG ; Chunyun LIU ; Weikun LI ; Chunyan MOU ; Li LIU
Journal of Clinical Hepatology 2025;41(8):1525-1532
Objective To investigate the predictive factors for the occurrence of HBsAg clearance in patients with HBeAg-negative chronic hepatitis B(CHB)receiving peginterferon alfa-2b(PEG-IFN-α-2b)treatment,analyze the effects of various indicators on the HBsAg clearance rate under different characteristics,and construct and evaluate a combined predictive model.Methods We included 125 patients with HBeAg-negative CHB at Kunming Third People's Hospital from May 2021 to May 2023.After treatment with PEG-IFN-α-2b combined with nucleoside analogues for a course of 48 weeks,they were divided into HBsAg clearance group and HBsAg non-clearance group.Their general information and serological,biochemical,and virological indicators at different time points during treatment were recorded.Continuous data in normal distribution were compared using the t test.Continuous data in non-normal distribution were compared using the Mann-Whitney U test,and comparisons across different time points were performed using the multiple paired-sample Friedman test.Categorical data were compared using the χ2 test.A Logistic regression analysis was used to select variables to establish a combined multi-parameter predictive model.Receiver operating characteristic(ROC)curves were generated to evaluate the diagnostic value of individual indicators and the combined predictive model for HBsAg clearance.Results Before treatment,there were significant differences in baseline HBsAg level(Z=-3.997,P<0.05)and treatment history(χ2=8.221,P<0.05)between the two groups.During treatment,gradually decreasing trends were observed in white blood cell count(χ2=104.944),neutrophil count(χ2=132.036),platelet count(χ2=162.881),and thyroid-stimulating hormone level(TSH,χ2=83.304,all P<0.05),while alanine aminotransferase(ALT,χ2=157.618)and alpha fetoprotein(χ2=159.472)showed gradually increasing trends(both P<0.05).At 48 weeks of treatment,treatment history(odds ratio[OR]=0.232,95%confidence interval[CI]:0.071-0.753),baseline HBsAg level(OR=13.423,95%CI:3.276-54.997),the extent of decrease in HBsAg from baseline after 12 weeks of treatment(OR=0.143,95%CI:0.040-0.515),the maximum ALT level during treatment(OR=0.986,95%CI:0.980-0.993),and the minimum TSH level during treatment(OR=3.281,95%CI:1.413-7.619)were independent factors affecting HBsAg clearance(all P<0.05).A combined predictive model for HBsAg clearance was built:Y=-1.603-1.462×treatment history+2.597×baseline HBsAg value-1.944×the extent of HBsAg reduction from baseline after 12 weeks of treatment-0.014×the maximum ALT value during treatment+1.188×the minimum TSH value during treatment.The diagnostic value of the individual indicators for HBsAg clearance from high to low was as following:the maximum ALT value during treatment(AUC=0.824),baseline HBsAg value(AUC=0.727),the minimum TSH value during treatment(AUC=0.707),the extent of HBsAg reduction from baseline after 12 weeks of treatment(AUC=0.641),and treatment history(AUC=0.636).The combined model showed better predictive performance than the individual indicators,with the AUC being 0.921(all P<0.05).Conclusion The combined model,constructed with baseline HBsAg value,the extent of HBsAg reduction from baseline after 12 weeks of treatment,the maximum ALT value during treatment,and the minimum TSH value during treatment,has high predictive value for the occurrence of HBsAg clearance in patients with HBeAg-negative CHB after 48 weeks of treatment with PEG-IFN-α-2b,which can provide a reference for identifying suitable patients for treatment and predicting clinical outcome.
6.Tubeimoside I promoted Snail ubiquitination degradation and inhibited the malignant progression of PANC-1 pancreatic cancer cells
Lixue FENG ; Chunyun ZHANG ; Zeyan LI ; Huiqi YIN ; Yingning SUN ; Dian-hui LIU ; Baogang YU ; He LIU ; Qingzhu YANG
Chinese Journal of Pathophysiology 2025;41(10):1955-1962
AIM:This study aims to investigate the molecular mechanism by which tubeimoside I(TBMS1)inhibits Snail expression in pancreatic cancer cells(PANC-1).METHODS:Human pancreatic cancer PANC-1 cells were cultured in vitro.The inhibitory effect of TBMS1 on PANC-1 cells was assessed using the MTT assay,and the data were analyzed based on the IC50 value of TBMS1.The impact of TBMS1 on the clonal formation ability of PANC-1 cells was evaluated through colony formation assays.The Transwell assay was employed to assess the effect of TBMS1 on the migrato-ry capability of PANC-1 cells.Apoptosis and cell cycle alterations in PANC-1 cells were analyzed using acridine orange staining and flow cytometry.The expression of Snail protein in pancreatic cancer and its relationship with survival of the patients were analyzed using the GEPIA database and Kaplan-Meier Plotter data.Immunofluorescence staining was con-ducted to investigate the effect of TBMS1 on Snail expression,while Western blot was used to evaluate the expression of poly(ADP-ribose)polymerase(PARP),E-cadherin and Snail in the cells.The ubiquitination of Snail protein was mea-sured using immunoprecipitation techniques.RESULTS:As the concentration of TBMS1 increased,the survival rate and number of clones formed by PANC-1 cells progressively decreased,leading to apoptosis,cleavage of PARP,and cell cycle arrest in the G1 phase.There was also a reduction in the proportion of cells in the S phase and a decrease in cell migration ability.The expression of Snail protein,a critical factor in cell migration,was inhibited,while E-cadherin protein levels were increased.Treatment with the proteasome inhibitor MG132 was able to reverse the suppression of Snail protein ex-pression caused by TBMS1.Immunoprecipitation results indicated that TBMS1 enhances the ubiquitination and subse-quent degradation of Snail protein.CONCLUSION:TBMS1 effectively inhibits the malignant progression of pancreatic cancer cells by promoting the ubiquitination and degradation of Snail protein in PANC-1 cells.
7.Comparison of two methods for establishing mouse models of acute exacer-bation of chronic obstructive pulmonary disease
Hongping ZHANG ; Runhua HOU ; Yuanyuan YE ; Zeping YANG ; Guanqin ZHENG ; Mengzhu LI ; Jiarui FAN ; Chunyun ZHANG ; Rui CHEN ; Defu LI
Chinese Journal of Pathophysiology 2025;41(10):2073-2080
AIM:To compare the degree of disease simulation between the two mouse models of acute exacer-bation of chronic obstructive pulmonary disease(AECOPD)using intranasal instillation of lipopolysaccharide(LPS)and fine particulate matter(PM2.5)for 3 d based on exposure to cigarette smoke(CS)for 90 d.METHODS:Thirty-two male C57BL/6 mice were randomly divided into 4 groups:control group(n=8),CS group(n=8),CS+PM2.5 group(n=8)and CS+LPS group(n=8).The AECOPD models in CS+PM2.5 and CS+LPS groups were established by CS exposure combined with intranasal PM2.5 and LPS instillation.Lung function,lung pathology and airway goblet cell hyperplasia using histologi-cal staining were measured.To evaluate the degree of lung inflammation and mucus secretion in mice,the prorein levels of mucin 5AC(MUC5AC),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the bronchoalveolar lavage fluid(BALF)were detected by ELISA and total white blood cell(WBC)counts and the BALF differential cell counts(neutro-phils,macrophages,lymphocytes)were detected by Giemsa staining.RESULTS:In CS group,lung function decreased(P<0.05),and bronchial inflammation index increased(P<0.01),airway goblet cell hyperplasia and airway collagen de-position were significant(P<0.01),total WBC count and differential cell count in the BALF increased(P<0.05),MUC5AC and inflammatory factor IL-6 and TNF-α levels increased(P<0.05),compared with control group.Compared with CS group,lung function decreased(P<0.05),the bronchial inflammation index increased(P<0.01),airway goblet cell hyperplasia and airway collagen deposition were significant(P<0.01),total WBC count and differential cell count in the BALF increased(P<0.05),and MUC5AC and inflammatory factor IL-6 and TNF-α levels increased(P<0.05)in CS+PM2.5 and CS+LPS groups.Compared with CS+PM2.5 group,lung function decreased(P<0.05),the bronchial inflamma-tion index increased(P<0.01),airway goblet cell hyperplasia and airway collagen deposition were significant(P<0.01),total WBC count and differential cell count in the BALF increased(P<0.05),and MUC5AC and inflammatory factor IL-6 and TNF-α levels increased(P<0.05)in CS+LPS group.CONCLUSION:Exposure to CS combined with both intrana-sal PM2.5 and LPS instillation allowed for establishing AECOPD models in mice,and CS exposure combined with intrana-sal LPS instillation better simulated AECOPD characteristics.
8.Sodium tanshinone ⅡA sulfonate inhalation attenuates pulmonary hyper-tension associated with chronic obstructive pulmonary disease in mice
Runhua HOU ; Siting WANG ; Hongping ZHANG ; Ruanyuan YE ; Zeping YANG ; Mengzhu LI ; Guanqing ZHEN ; Jiarui FAN ; Chunyun ZHANG ; Rui CHEN ; Defu LI
Chinese Journal of Pathophysiology 2025;41(9):1713-1720
AIM:To investigate the effects of nebulized sodium tanshinone ⅡA sulfonate(STS)in a mouse model of pulmonary hypertension associated with chronic obstructive pulmonary disease(COPD-PH).METHODS:A to-tal of 32 healthy SPF-grade male C57BL/6 mice were randomly divided into 4 groups:control(CTL,n=8)group,COPD-PH(CS+LPS,n=8)group,STS-treated COPD-PH(CS+LPS+STS,n=8)group,and STS(n=8)group.The COPD-PH model was established through whole-body exposure to cigarette smoke(CS)combined with lipopolysaccharide(LPS)in-halation.Mice were subjected to cigarette smoke exposure in a chamber(9 cigarettes/h,2 h/session,2 sessions/d,6 d/week)for 60 d,except on days of LPS inhalation.On days 1 and 14,COPD-PH model mice received LPS(7.5 μg/mouse in 50 μL saline)via intranasal inhalation,while the CTL and STS groups received an equivalent volume of saline.STS was administered via nebulized inhalation(5 mg/kg,30 min per session,twice daily)immediately before CS exposure.At the end of the modeling period,lung function and right heart pressure were assessed.Bronchoalveolar lavage fluid(BALF)was collected for inflammatory cell counting.Levels of interleukin-6(IL-6)in BALF supernatants and plasma were measured using ELISA.Pathological changes in the airway and lung tissues were evaluated.RESULTS:(1)Com-pared to CTL mice,those exposed to CS and LPS exhibited lesions characteristic of COPD-PH,including emphysema,lung inflammation,decreased lung function,and increased right ventricular systolic pressure(RVSP)and right ventricu-lar hypertrophy index(RVHI)(P<0.05);(2)COPD-PH mice showed significantly elevated IL-6 levels in both BALF and plasma(P<0.05);(3)STS treatment alleviated emphysema and lung inflammation,improved lung function,prevent-ed increases in RVSP and the RV/(LV+S)ratio,and reduced IL-6 levels in both BALF and plasma(P<0.05).CON-CLUSION:The results indicate that nebulized inhalation of STS significantly slows the progression of COPD-PH,likely due to its ability to inhibit lung inflammation and reduce IL-6 expression in the lungs.
9.Tubeimoside I promoted Snail ubiquitination degradation and inhibited the malignant progression of PANC-1 pancreatic cancer cells
Lixue FENG ; Chunyun ZHANG ; Zeyan LI ; Huiqi YIN ; Yingning SUN ; Dian-hui LIU ; Baogang YU ; He LIU ; Qingzhu YANG
Chinese Journal of Pathophysiology 2025;41(10):1955-1962
AIM:This study aims to investigate the molecular mechanism by which tubeimoside I(TBMS1)inhibits Snail expression in pancreatic cancer cells(PANC-1).METHODS:Human pancreatic cancer PANC-1 cells were cultured in vitro.The inhibitory effect of TBMS1 on PANC-1 cells was assessed using the MTT assay,and the data were analyzed based on the IC50 value of TBMS1.The impact of TBMS1 on the clonal formation ability of PANC-1 cells was evaluated through colony formation assays.The Transwell assay was employed to assess the effect of TBMS1 on the migrato-ry capability of PANC-1 cells.Apoptosis and cell cycle alterations in PANC-1 cells were analyzed using acridine orange staining and flow cytometry.The expression of Snail protein in pancreatic cancer and its relationship with survival of the patients were analyzed using the GEPIA database and Kaplan-Meier Plotter data.Immunofluorescence staining was con-ducted to investigate the effect of TBMS1 on Snail expression,while Western blot was used to evaluate the expression of poly(ADP-ribose)polymerase(PARP),E-cadherin and Snail in the cells.The ubiquitination of Snail protein was mea-sured using immunoprecipitation techniques.RESULTS:As the concentration of TBMS1 increased,the survival rate and number of clones formed by PANC-1 cells progressively decreased,leading to apoptosis,cleavage of PARP,and cell cycle arrest in the G1 phase.There was also a reduction in the proportion of cells in the S phase and a decrease in cell migration ability.The expression of Snail protein,a critical factor in cell migration,was inhibited,while E-cadherin protein levels were increased.Treatment with the proteasome inhibitor MG132 was able to reverse the suppression of Snail protein ex-pression caused by TBMS1.Immunoprecipitation results indicated that TBMS1 enhances the ubiquitination and subse-quent degradation of Snail protein.CONCLUSION:TBMS1 effectively inhibits the malignant progression of pancreatic cancer cells by promoting the ubiquitination and degradation of Snail protein in PANC-1 cells.
10.Comparison of two methods for establishing mouse models of acute exacer-bation of chronic obstructive pulmonary disease
Hongping ZHANG ; Runhua HOU ; Yuanyuan YE ; Zeping YANG ; Guanqin ZHENG ; Mengzhu LI ; Jiarui FAN ; Chunyun ZHANG ; Rui CHEN ; Defu LI
Chinese Journal of Pathophysiology 2025;41(10):2073-2080
AIM:To compare the degree of disease simulation between the two mouse models of acute exacer-bation of chronic obstructive pulmonary disease(AECOPD)using intranasal instillation of lipopolysaccharide(LPS)and fine particulate matter(PM2.5)for 3 d based on exposure to cigarette smoke(CS)for 90 d.METHODS:Thirty-two male C57BL/6 mice were randomly divided into 4 groups:control group(n=8),CS group(n=8),CS+PM2.5 group(n=8)and CS+LPS group(n=8).The AECOPD models in CS+PM2.5 and CS+LPS groups were established by CS exposure combined with intranasal PM2.5 and LPS instillation.Lung function,lung pathology and airway goblet cell hyperplasia using histologi-cal staining were measured.To evaluate the degree of lung inflammation and mucus secretion in mice,the prorein levels of mucin 5AC(MUC5AC),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the bronchoalveolar lavage fluid(BALF)were detected by ELISA and total white blood cell(WBC)counts and the BALF differential cell counts(neutro-phils,macrophages,lymphocytes)were detected by Giemsa staining.RESULTS:In CS group,lung function decreased(P<0.05),and bronchial inflammation index increased(P<0.01),airway goblet cell hyperplasia and airway collagen de-position were significant(P<0.01),total WBC count and differential cell count in the BALF increased(P<0.05),MUC5AC and inflammatory factor IL-6 and TNF-α levels increased(P<0.05),compared with control group.Compared with CS group,lung function decreased(P<0.05),the bronchial inflammation index increased(P<0.01),airway goblet cell hyperplasia and airway collagen deposition were significant(P<0.01),total WBC count and differential cell count in the BALF increased(P<0.05),and MUC5AC and inflammatory factor IL-6 and TNF-α levels increased(P<0.05)in CS+PM2.5 and CS+LPS groups.Compared with CS+PM2.5 group,lung function decreased(P<0.05),the bronchial inflamma-tion index increased(P<0.01),airway goblet cell hyperplasia and airway collagen deposition were significant(P<0.01),total WBC count and differential cell count in the BALF increased(P<0.05),and MUC5AC and inflammatory factor IL-6 and TNF-α levels increased(P<0.05)in CS+LPS group.CONCLUSION:Exposure to CS combined with both intrana-sal PM2.5 and LPS instillation allowed for establishing AECOPD models in mice,and CS exposure combined with intrana-sal LPS instillation better simulated AECOPD characteristics.

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