1.Real-world efficacy and safety of azvudine in hospitalized older patients with COVID-19 during the omicron wave in China: A retrospective cohort study.
Yuanchao ZHU ; Fei ZHAO ; Yubing ZHU ; Xingang LI ; Deshi DONG ; Bolin ZHU ; Jianchun LI ; Xin HU ; Zinan ZHAO ; Wenfeng XU ; Yang JV ; Dandan WANG ; Yingming ZHENG ; Yiwen DONG ; Lu LI ; Shilei YANG ; Zhiyuan TENG ; Ling LU ; Jingwei ZHU ; Linzhe DU ; Yunxin LIU ; Lechuan JIA ; Qiujv ZHANG ; Hui MA ; Ana ZHAO ; Hongliu JIANG ; Xin XU ; Jinli WANG ; Xuping QIAN ; Wei ZHANG ; Tingting ZHENG ; Chunxia YANG ; Xuguang CHEN ; Kun LIU ; Huanhuan JIANG ; Dongxiang QU ; Jia SONG ; Hua CHENG ; Wenfang SUN ; Hanqiu ZHAN ; Xiao LI ; Yafeng WANG ; Aixia WANG ; Li LIU ; Lihua YANG ; Nan ZHANG ; Shumin CHEN ; Jingjing MA ; Wei LIU ; Xiaoxiang DU ; Meiqin ZHENG ; Liyan WAN ; Guangqing DU ; Hangmei LIU ; Pengfei JIN
Acta Pharmaceutica Sinica B 2025;15(1):123-132
Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T NANC), time to symptom improvement (T SI), and time of hospital stay (T HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.
2.Effect of umbilical cord mesenchymal stem cell-derived exosomes on oxidative damage and expressions of inflammatory factors in endometrial stromal cells
Lin WANG ; Xuguang MI ; Xiuying LIN ; Jianhua FU ; Lei LIU ; Aiqiao WANG ; Qian DU ; Wenqi ZHANG ; Meijiao FAN ; Yanqiu FANG
Chinese Journal of Immunology 2025;41(9):2153-2160
Objective:To investigate effects of human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSCs-Exo)on bisphenol AF(BPAF)-induced oxidative damage and inflammatory factor release from endometrial stromal cells(hESCs).Methods:hESCs were divided into Control group,BPAF group(25 μmol/L BPAF treatment),BPAF+Exo group(25 μmol/L BPAF+hUCMSCs-Exo treatment),BPAF+Exo+LY group(25 μmol/L BPAF+hUCMSCs-Exo+10 μmol/L LY294002 treatment).Cell prolifera-tion was detected by MTT assay;apoptosis,intracellular ROS level,and mitochondrial membrane potential level were detected by flow cytometry;protein expressions of Bcl-2,Bax,Cleaved-caspase-3 and PI3K/AKT signaling pathway were detected by Western blot;mRNA expressions of inflammatory factors TNF-α,IL-6 and IL-1β were detected by RT-qPCR.Results:Compared with Control group,hESCs survival rate was gradually decreased(P<0.01),apoptosis rate was gradually increased with the increased concentration of BPAF(≥25 μmol/L).Compared with Control group,BPAF group showed increased ROS level,decreased mitochondrial membrane potential level,increased Bax and Cleaved-caspase-3 protein expressions,and decreased Bcl-2,p-PI3K and p-AKT protein expressions.Compared with BPAF group,cell survival rate of BPAF+Exo group was increased(P<0.01),ROS level decreased,mitochondrial membrane potential level increased,expressions of Bax and Cleaved-caspase-3 proteins decreased,and expressions of Bcl-2,p-PI3K and p-AKT increased.Compared with BPAF+Exo group,expressions of Bax and Cleaved-caspase-3 protein in cells of BPAF+Exo+LY group were increased,while expressions of Bcl-2,p-PI3K and p-AKT protein were decreased.Expressions of inflammatory factors TNF-α,IL-6 and IL-1β mRNA were significantly up-regulated in BPAF group compared with Control group(P<0.01),and expressions of inflammatory factors mRNA were significantly down-regulated in BPAF+Exo group compared with BPAF group(P<0.05).Conclu-sion:BPAF(≥25 μmol/L)inhibits proliferation of hESCs and promoted apoptosis.hUCMSCs-Exo inhibits BPAF-induced oxidative dam-age and inflammatory factors expressions in hESCs through PI3K/AKT signaling pathway.
3.Epidemic characteristics and healthcare-seeking behavior of dengue fever cases in Zhejiang Province in 2024
Song GUO ; Ying LIU ; Jiangping REN ; Rong ZHANG ; Xuguang SHI ; Jimin SUN
Chinese Journal of Endemiology 2025;44(10):831-835
Objective:To study the epidemic characteristics and healthcare-seeking behavior of dengue fever cases in Zhejiang Province, and to provide a scientific basis for developing dengue fever prevention and control strategies.Methods:Data on dengue fever epidemic in Zhejiang Province from January to December 2024 were collected from the Information System for China Disease Control and Prevention and individual case reports. Descriptive epidemiological methods were used to analyze the distribution by the three dimensions (time, region, and population), healthcare-seeking behavior, importation sources, and laboratory test results of dengue fever cases in Zhejiang Province.Results:From January to December 2024, Zhejiang Province reported a total of 289 dengue fever cases (including 25 foreigners), with an incidence rate of 0.44/100 000, including one severe case and no death, and 13 were local, 108 were domestically imported, and 168 were internationally imported. The epidemic had affected 70 districts (counties, cities) across all 11 cities in the province. Cases were reported monthly, with the peak incidence concentrated from July to November. The age of dengue fever cases was 38 (30, 48) years old, ranging from 2 to 75 years old, with a higher incidence in the 30 - 39 age group (31.14%, 90/289). The male-to-female ratio was 1.86 ∶ 1.00 (188 ∶ 101). Occupational distribution was predominantly commercial service workers (28.03%, 81/289). The number of healthcare-seeking for local cases was 2 (1, 2) times, ranging from 1 to 6 times. The number of healthcare-seeking for imported cases was 1 (1, 2) times, ranging from 1 to 4 times. The interval between the onset and diagnosis of local cases was 3.5 (1.0, 5.0) days, ranging from 0 to 6 days. The interval between the onset and diagnosis of imported cases was 3(1, 4) days, ranging from 0 to 6 days. Domestically imported cases originated from Guangdong Province (102 cases), Yunnan Province (3 cases), Hunan Province (1 case), with 2 cases had unknown original provinces. Internationally imported cases mainly came from Indonesia (33 cases), Thailand (22 cases), Laos (18 cases). Serotyping of dengue virus was determined for 283 cases, with 157 cases of type Ⅰ, 88 cases of type Ⅱ, 34 cases of type Ⅲ, and 4 cases of type Ⅳ.Conclusions:The dengue fever cases in Zhejiang Province in 2024 is primarily international importation, the interval between the onset and diagnosis of local cases is relatively long. Prevention and control efforts should strengthen surveillance and early warning based on dengue fever's epidemic characteristics, and reinforcing port quarantine, mosquito vector control, and health education for high-risk populations.
4.Effect of umbilical cord mesenchymal stem cell-derived exosomes on oxidative damage and expressions of inflammatory factors in endometrial stromal cells
Lin WANG ; Xuguang MI ; Xiuying LIN ; Jianhua FU ; Lei LIU ; Aiqiao WANG ; Qian DU ; Wenqi ZHANG ; Meijiao FAN ; Yanqiu FANG
Chinese Journal of Immunology 2025;41(9):2153-2160
Objective:To investigate effects of human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSCs-Exo)on bisphenol AF(BPAF)-induced oxidative damage and inflammatory factor release from endometrial stromal cells(hESCs).Methods:hESCs were divided into Control group,BPAF group(25 μmol/L BPAF treatment),BPAF+Exo group(25 μmol/L BPAF+hUCMSCs-Exo treatment),BPAF+Exo+LY group(25 μmol/L BPAF+hUCMSCs-Exo+10 μmol/L LY294002 treatment).Cell prolifera-tion was detected by MTT assay;apoptosis,intracellular ROS level,and mitochondrial membrane potential level were detected by flow cytometry;protein expressions of Bcl-2,Bax,Cleaved-caspase-3 and PI3K/AKT signaling pathway were detected by Western blot;mRNA expressions of inflammatory factors TNF-α,IL-6 and IL-1β were detected by RT-qPCR.Results:Compared with Control group,hESCs survival rate was gradually decreased(P<0.01),apoptosis rate was gradually increased with the increased concentration of BPAF(≥25 μmol/L).Compared with Control group,BPAF group showed increased ROS level,decreased mitochondrial membrane potential level,increased Bax and Cleaved-caspase-3 protein expressions,and decreased Bcl-2,p-PI3K and p-AKT protein expressions.Compared with BPAF group,cell survival rate of BPAF+Exo group was increased(P<0.01),ROS level decreased,mitochondrial membrane potential level increased,expressions of Bax and Cleaved-caspase-3 proteins decreased,and expressions of Bcl-2,p-PI3K and p-AKT increased.Compared with BPAF+Exo group,expressions of Bax and Cleaved-caspase-3 protein in cells of BPAF+Exo+LY group were increased,while expressions of Bcl-2,p-PI3K and p-AKT protein were decreased.Expressions of inflammatory factors TNF-α,IL-6 and IL-1β mRNA were significantly up-regulated in BPAF group compared with Control group(P<0.01),and expressions of inflammatory factors mRNA were significantly down-regulated in BPAF+Exo group compared with BPAF group(P<0.05).Conclu-sion:BPAF(≥25 μmol/L)inhibits proliferation of hESCs and promoted apoptosis.hUCMSCs-Exo inhibits BPAF-induced oxidative dam-age and inflammatory factors expressions in hESCs through PI3K/AKT signaling pathway.
5.Epidemic characteristics and healthcare-seeking behavior of dengue fever cases in Zhejiang Province in 2024
Song GUO ; Ying LIU ; Jiangping REN ; Rong ZHANG ; Xuguang SHI ; Jimin SUN
Chinese Journal of Endemiology 2025;44(10):831-835
Objective:To study the epidemic characteristics and healthcare-seeking behavior of dengue fever cases in Zhejiang Province, and to provide a scientific basis for developing dengue fever prevention and control strategies.Methods:Data on dengue fever epidemic in Zhejiang Province from January to December 2024 were collected from the Information System for China Disease Control and Prevention and individual case reports. Descriptive epidemiological methods were used to analyze the distribution by the three dimensions (time, region, and population), healthcare-seeking behavior, importation sources, and laboratory test results of dengue fever cases in Zhejiang Province.Results:From January to December 2024, Zhejiang Province reported a total of 289 dengue fever cases (including 25 foreigners), with an incidence rate of 0.44/100 000, including one severe case and no death, and 13 were local, 108 were domestically imported, and 168 were internationally imported. The epidemic had affected 70 districts (counties, cities) across all 11 cities in the province. Cases were reported monthly, with the peak incidence concentrated from July to November. The age of dengue fever cases was 38 (30, 48) years old, ranging from 2 to 75 years old, with a higher incidence in the 30 - 39 age group (31.14%, 90/289). The male-to-female ratio was 1.86 ∶ 1.00 (188 ∶ 101). Occupational distribution was predominantly commercial service workers (28.03%, 81/289). The number of healthcare-seeking for local cases was 2 (1, 2) times, ranging from 1 to 6 times. The number of healthcare-seeking for imported cases was 1 (1, 2) times, ranging from 1 to 4 times. The interval between the onset and diagnosis of local cases was 3.5 (1.0, 5.0) days, ranging from 0 to 6 days. The interval between the onset and diagnosis of imported cases was 3(1, 4) days, ranging from 0 to 6 days. Domestically imported cases originated from Guangdong Province (102 cases), Yunnan Province (3 cases), Hunan Province (1 case), with 2 cases had unknown original provinces. Internationally imported cases mainly came from Indonesia (33 cases), Thailand (22 cases), Laos (18 cases). Serotyping of dengue virus was determined for 283 cases, with 157 cases of type Ⅰ, 88 cases of type Ⅱ, 34 cases of type Ⅲ, and 4 cases of type Ⅳ.Conclusions:The dengue fever cases in Zhejiang Province in 2024 is primarily international importation, the interval between the onset and diagnosis of local cases is relatively long. Prevention and control efforts should strengthen surveillance and early warning based on dengue fever's epidemic characteristics, and reinforcing port quarantine, mosquito vector control, and health education for high-risk populations.
6.Surveillance and analysis of leptospirosis in Zhejiang Province in 2023
Song GUO ; Wenwu YAO ; Ying LIU ; Xuguang SHI ; Jiangping REN ; Rong ZHANG ; Mingyu LUO ; Zhangnyu YANG ; Jimin SUN
Chinese Journal of Endemiology 2024;43(7):559-563
Objective:The epidemiological and host animal pathogen data of leptospirosis in the population of Zhejiang Province in 2023 were analyzed, providing scientific basis for formulating prevention and control strategies of leptospirosis.Methods:The data of human leptospirosis in the population were collected from the China Information System for Disease Control and Prevention, and analyzed using descriptive epidemiological methods. The data on isolation, culture, and nucleic acid testing of Leptospira pathogens from mouse kidneys, frog kidneys, pig kidneys, and duck kidneys as well as duck serum antibody data were collected from Zhejiang Provincial Center for Disease Control and Prevention "Leptospirosis Surveillance Project of Zhejiang Province". The carrying and changing status of Leptospira epidemic microbiota in populations and host animals were analyzed. Results:In 2023, a total of 83 cases of leptospirosis were reported in Zhejiang Province, with a incidence rate of 0.126 2/ 100 000, aged (62.66 ± 11.31) years, including 68 males and 15 females. Leptospirosis cases were reported in 11 cities, mainly concentrated in the southern cities of Wenzhou City, Lishui City and Taizhou City(a total of 68 cases), accounting for 81.93% of the total number of cases. August to October were high incidence months for leptospirosis (a total of 70 cases), accounting for 84.34% of the total number cases. The male to female ratio of patients was 4.53 ∶ 1.00, and all were adults ≥20 years old, the middle and old people aged 45 - 79 years were the high-risk population (a total of 77 cases), accounting for 92.77% of the total number of cases. The patient's occupation was mainly farmers, with a total of 54 cases, accounting for 65.06% of the total number cases. The shortest time from onset to initial diagnosis for patients with leptospirosis was 0 day, and the longest was 13 days. The shortest time from initial diagnosis to confirmed diagnosis was 0 day, and the longest was 16 days. The 72.29% of the leptospirosis cases (60 cases) had a history of field labor or suspected contact with contaminated water within one month before the onset of the disease, and 18.07% of the leptospirosis cases (15 cases) had a history of contact with animals such as mice, frogs, pigs, cows, dogs, fish or ducks, or their excreta within one month before the onset of the disease. The average nucleic acid positive rate of host animals with leptospirosis was 5.92% (31/524) in mouse kidney, 6.74% (36/534) in frog kidney, and 0.66% (1/151) in pig kidney. The isolation and culture of leptospirosis from duck kidney, nucleic acid detection, and antibody detection in duck blood were all negative. The leptospirosis bacteria detected in human population were serogroup Icterohaemorrhagiae (3 samples) and Hebdomadis (4 samples), and the bacteria group detected in host animals was serogroup Icterohaemorrhagiae (3 samples). Conclusions:The outbreak of leptospirosis in Zhejiang Province mainly occurs in the summer and autumn, with the affected areas mainly concentrated in the southern region of Zhejiang Province. The affected population is mainly middle-aged and elderly males, and the population carrying Leptospira is still mainly composed of the serogroup Icterohaemorrhagiae and the Hebdomadis, with the host animal being the serogroup Icterohaemorrhagiae.
7.Effect of spermidine on autophagy and inflammatory cytokine expression in human endometrial stromal cells
Xue ZHANG ; Xuguang MI ; Xiuying LIN ; Jianhua FU ; Lei LIU ; Xinyue GAO ; Yanqiu FANG
Chinese Journal of Immunology 2024;40(5):1023-1029
Objective:To investigate damaging effects of clomifene citrate(CC)on endometrial stromal cells(hEndoSCs),and to study effects of spermidine on autophagy and inflammatory cytokine expression in damaged endometrial stromal cells.Methods:Groups were firstly divided into control group,spermidine group,clomiphene group(CC group),CC+Spermidine group.MTT assay was used to detect cell survival rate of hEndoSCs after co-incubation with different concentrations of CC or Spermidine for 24 h.Con-tent of intracellular reactive oxygen species(ROS)and level of apoptosis in cells of the 4 groups were detected by flow cytometry tech-nique.Western blot was used to detect expressions of autophagy pathway-related proteins ULK1,p-ULK1,LC-3Ⅱ,and apoptosis-re-lated proteins Bax,Bcl-2,Cleaved-caspase 3.RT-qPCR was used to detect mRNA expressions of IL-6,IL-1β and TNF-α.Results:Compared with control group,CC group showed decreased cell survival,increased apoptosis rate,ROS content,Bax,Cleaved-cas-pase 3 expressions,decreased Bcl-2 expression,decreased levels of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ,and elevated expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA(P<0.01).There was no significant changes viability of cells in spermidine group compared with control group(P>0.05).Compared with CC group,cell survival rate in CC+spermidine group was sig-nificantly increased,apoptosis rate,ROS content,Bax and Cleaved-caspase 3 expressions were decreased,Bcl-2 expression was in-creased,expressions of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ were elevated,while expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA were decreased(P<0.01).Conclusion:CC can inhibit endometrial stromal cell proliferation,promote apoptosis,and increase the transcript levels of inflammatory factors IL-6,IL-1β and TNF-α.Spermidine can reduce intracellular ROS in clomiphene-injured endometrial stromal cells by activating cellular autophagy,increase cell survival,and inhibit the expressions of inflammatory factors IL-6,IL-1β and TNF-α.
8.Effect of Wnt/β-catenin signaling pathway inhibitor MS AB on fibrogenic responses of human endometrial stromal cells
Feina WANG ; Xuguang MI ; Xiuying LIN ; Jianhua FU ; Lei LIU ; Xinyue YU ; Huanhuan ZANG ; Linjun LIU ; Shiling CHEN ; Yanqiu FANG
Journal of Jilin University(Medicine Edition) 2024;50(5):1266-1274
Objective:To discuss the effect of Wnt/β-catenin signaling pathway inhibitor methyl 3-{[(4-methyl-phenyl)sulfonyl]amino } benzoate(MS AB)on the fibrogenic response of the human endometrial stromal cells(HESCs),and to provide the foundation for the application of MSAB in the target therapy of intrauteriue adhesion(JUA).Methods:The normal HESCs were cultured in vitro and divided into two groups:control group and transforming growth factor β1(TGF-β1)group;the HESCs from the adhesion part of the IUA patients were cultured in vitro,regarded as IUA group.Western blotting method was used to detect the expression levels of fibrotic marker protein type Ⅰ collagen α1(COL1A1)in the cells in various groups at different time points(0,12,24,48,and 60 h)after treated with TGF-β1.MTT assay was used to detect the proliferation activities of the cells in various groups.Western blotting method was used to detect the expression levels of the fibrotic marker protein COL1A1,stromal marker proteins such as N-cadherin and α-smooth muscle actin(α-SMA),and Wnt/β-catenin signaling pathway-related protein β-catenin in the cells in control and IUA groups.Based on the MSAB concentrations,the normal HESCs were divided into 0(control),0.25,0.50,0.75,and 1.00 μmol·L-1 MSAB groups,and MTT assay was used to detect the survival rates of the cells in various groups.After treated with MSAB,the normal HESCs were divided into control group(normal HESCs),TGF-β1 group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with complete culture medium,and the cells continued to be cultured for 24 h),and MSAB group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with a complete medium containing 0.75 μmol·L-1 MSAB and the cells continued to be cultured for 24 h).Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of epithelial-mesenchymal transition(EMT)-related transcription factors Snail,Slug,Smuc,ZEB1,and ZEB2,and COL1A1 mRNA in the cells in various groups.Western blotting method was used to detect the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in various groups.Results:Compared with control group(after treated with TGF-β1 for 0 h),the expression levels of COL1A1 proteins in the HESCs after treated with TGF-β1 for 12,24,48,and 60 h in TGF-β1 group were increased(P<0.05 or P<0.01).Compared with control group,there was no significant difference in the proliferation activity of the HESCs in IUA and TGF-β1 groups(P>0.05).Compared with control group,the expression levels of COL1A1,β-catenin,N-cadherin,and α-SMA proteins in the cells in IUA group were increased(P<0.05 or P<0.01).Compared with control group,the survival rates of the cells in 0.75 and 1.00 μmol·L-1 MSAB groups were decreased(P<0.05 or P<0.01).Compared with control group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in TGF-β1 group were increased(P<0.05 or P<0.01);compared with TGF-β1 group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in MSAB group were decreased(P<0.05 or P<0.01).Compared with control group,after treated with TGF-β1 for 24 h,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in TGF-β1 group were increased(P<0.01);compared with TGF-β1 group,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in MSAB group were decreased(P<0.05 or P<0.01).Conclusion:MSAB can inhibit the fibrogenic responses of the HESCs in vitro,and the results provide the theoretical basis for the application of MSAB in the target therapy of IUA.
9.Effects of bisphenol A on stemness of endometrial mesenchymal stem/stromal cells and improvement effect of human umbilical cord mesenchymal stem cell-derived supernatant on cell injury
Aiqiao WANG ; Xuguang MI ; Xiuying LIN ; Jianhua FU ; Lei LIU ; Lin WANG ; Wenqi ZHANG ; Ling DENG ; Shiling CHEN ; Yanqiu FANG
Journal of Jilin University(Medicine Edition) 2024;50(6):1557-1564
Objective:To investigate the effects of bisphenol A(BPA)on the proliferation activity and stemness characteristics of endometrial mesenchymal stem/stromal cells(eMSCs),and to elucidate the improvement effect of human umbilical cord mesenchymal stem cell-derived supernatant(hUCMSC-Sup)on the cell injury.Methods:The eMSCs were cultured in vitro and treated with different concentrations of BPA(0,200,250,300,350,and 400 μmol·L-1).The eMSCs were divided into control group(only cultured with culture solution),BPA group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA),BPA+hUCMSC-Sup group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 50%volumetric ratio of hUCMSC-Sup),and BPA+CHIR-99021 group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 10 μmol·L-1 CHIR-99021).The survival rates of eMSCs in various groups were detected by methyl thiazolyl tetrazolium(MTT)assay.The numbers and diameters of the spheroids in various groups were detected by spheroids formation assay,the proliferation activities of the cells in eMSCs stem cell spheroids in various groups were detected by CCK-8 assay;the percentage of CD73+cells in eMSCs in various groups were detected by flow cytometry;the expression levels of sex determining region Y-box 2(Sox2),octamer-binding transcription factor 4(Oct4),and Nanog mRNA in the eMSCs in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method,the expression levels of β-catenin protein in the eMSCs in various groups were detected by Western blotting method.Results:The MTT results showed that after treated with BPA for 24 and 48 h,compared with 0 μmol·L-1 BPA group,the survival rates of eMSCs in 200,250,300,350,and 400 μmol·L-1 BPA groups were significantly decreased(P<0.01).At 24 and 48 h after treatment,compared with control group,the survival rate of the eMSCs in BPA group was significantly decreased(P<0.01);at 48 h after treatment,compared with BPA group,the survival rate of the eMSCs in BPA+hUCMSC-Sup group was significantly inereased(P<0.05).The spheroids formation assay results showed that compared with culture 3 d group,the numbers and diameters of stem cell spheroids of the eMSCs in culture 4 d group and culture 5 d group were significantly increased(P<0.05 or P<0.01);compared with control group,after 48 h of culture,the number and diameter of the cells in eMSCs stem cell spheroids in BPA group were significantly decreased(P<0.05 or P<0.01).The CCK-8 results showed that after 24 and 48 h of treatment,compared with control group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA group was significantly decreased(P<0.01);compared with BPA group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The flow cytometry results showed that compared with control group,the percentage of the CD73+cells in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the percentage of the CD73+cells in eMSCs in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The RT-qPCR results showed that compared with control group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA group were significantly decreased(P<0.01);compared with BPA group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression level of β-catenin protein in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the expression levels of β-catenin protein in the eMSCs in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were signifrcantly inereased(P<0.01).Conclusion:BPA can inhibit the stemness characteristics of the eMSCs,and injury the self-renewal and repair of endometrium;its mechanism may be related to down-regulating the activity of Wnt/β-catenin signal pathway in the cells.hUCMSC-Sup can promote the proliferation of injured eMSCs,and has improvement effect on the stemness injury induced by BPA.
10.Tea polyphenols ameliorates acute lung injury in septic mice by inhibiting NLRP3 inflammasomes
Xuguang LING ; Wenwen XU ; Guanlai PANG ; Xuxing HONG ; Fengqin LIU ; Yang LI
Journal of Southern Medical University 2024;44(2):381-386
Objective To investigate the mechanism of tea polyphenols(TP)for regulating NLRP3 inflammasomes and alleviating acute lung injury in septic mice.Methods Sixty C57BL/6 mice were randomly assigned into sham-operated,cecal ligation and puncture(CLP)and CLP+TP treatment groups,and survival of the mice was recorded after modeling in each group.The lung wet/dry weight ratio and myeloperoxidase(MPO)activity were determined,and lung injury of the mice was evaluated using HE staining and acute lung injury score.The expressions of IL-1β,TNF-α,IL-6,NLRP3,caspase-1 p10,ASC,MPO,and caspase-8 in the lung tissue were detected using ELISA,Western blotting,or immunohistochemical staining.MDA and H2O2 levels in the lungs were detected to evaluate the level of oxidative stress.Immunofluorescence assay was used to investigate the co-localization of NLRP3 and NOX4.Results The postoperative mortality rate at 72 h,lung wet/dry weight ratio,MPO level and acute lung injury scores were significantly lower in CLP+TP group than in CLP group(P<0.05).Treatment with TP significantly reduced the expressions of NLRP3-related inflammatory factors(P<0.05)and lowered MDA and H2O2 levels in the lung tissue of the septic mice(P<0.05).Immunofluorescence co-staining showed a lower level of NOX4 and NLRP3 co-localization in CLP+TP group than in CLP group.Conclusion TP inhibits NLRP3 inflammasome-associated inflammation to alleviate CLP-induced acute lung injury in mice through a regulatory mechanism that inhibits NOX4 expression and reduces oxidative stress in the lung tissue.

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