1.Study on the synergistic effects and mechanisms of IL-25 and house dust mite in promoting eosinophilic asthma allergy
Lina Xu ; Xiaoshuang He ; Dong Liu
Acta Universitatis Medicinalis Anhui 2025;60(4):649-655
Objective :
To explore the central role of interleukin(IL)-25 in allergy of eosinophil asthma.
Methods :
Forty 4-5 week-old C57BL/6 mice were used in this study(n=10). Mice were randomly divided into 4 groups: WT+shRNA NT group, WT+HDM group, IL-25 shRNA+IL-25 group, and IL-25 shRNA+IL-25+HDM group. According to each group′s treatment requirement, mice were treated with house dust mite and/or IL-25 by intranasal infusion. HE staining and PAS staining were used for lung tissue section staining and pathological analysis. The levels of type 2 cytokines in bronchoalveolar lavage fluid(BALF) were determined by ELISA. Flow cytometry(FCM) was used to determine the number of type 2 helper T(Th2) cells, the group 2 innate lymphoid cells(ILC2) and eosinophils in lung tissues. Mice were treated with FITC-labeled DQ-ovalbumin(DQ-OVA) by intranasal infusion to determine the antigen presentation ability of eosinophils infiltrated in the lungs.
Results :
HE staining and PAS staining results showed that, compared with WT+shRNA NT group, a large amount of mucus and a large number of eosinophil infiltration were found in the lungs, and subcutaneous thickening of inflammatory airway were observed in the pulmonary vessels, alveolar ducts and the whole alveoli in IL-25 shRNA+IL-25+HDM group. In the IL-25 shRNA+IL-25 group, only a small amount of mucus and a small amount of eosinophil infiltration were found in the lungs, and a small amount of subcutaneous thickening of inflammatory airway were observed in pulmonary vessels, alveolar ducts and the whole alveoli. There was no significant difference in WT+HDM group. Compared with WT+shRNA NT group, the levels of IL-25, IL-4,IL-5,IL-13, IL-33 and interferon-γ(IFN-γ) in BALF significantly increased; the levels of infiltrated Th2, ILC2 and eosinophils in the lung significantly increased; the fluorescence levels of FITC-labeled DQ-OVA in eosinophils infiltrated in lungs significantly increased in IL-25 shRNA+IL-25+HDM group(P<0.05). IL-25 shRNA+IL-25 group mice also showed an increase in the above measurements, but the amplitude were limited(P<0.05). WT+HDM group had no significant difference.
Conclusion
IL-25 plays an essential role in the allergy and eosinophil antigen presentation processes in eosinophil asthma mice.
2.Effects of lncRNA DUXAP8 in lung cancer A549 cells-derived exosomes on lung cancer cell growth and its mechnism
Xiaoshuang HE ; Lina XU ; Mei CUI ; Yu ZHAO ; Bei WANG ; Zheng HUANG ; Yuchao WANG ; Wenyan XIN ; Chao WU
Journal of Jilin University(Medicine Edition) 2025;51(4):958-967
Objective:To discuss the effect of long non-coding RNA(lncRNA)DUXAP8 in exosomes(Exo)derived from the lung cancer A549 cells on the growth and immune escape of the lung cancer cells,and to clarify the mechanism.Methods:The human lung cancer cell line A549 was cultured,and its exosomes were extracted and identified.The A549 cells were treated with PKH67-labeled Exo to observe the uptake of Exo by A549 cells.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression level of lncRNA DUXAP8 in A549 cells before and after Exo treatment.The A549 cells were divided into control group(no treatment),Exo group(A549 cells treated with Exo),Exo+sh-NC group(A549 cells treated with Exo and then transfected with sh-NC),and Exo+sh-DUXAP8 group(A549 cells treated with Exo and then transfected with sh-DUXAP8).RT-qPCR method was used to detect the expression level of lncRNA DUXAP8 in A549 cells in various groups;colony formation assay was used to detect the colony formation abilities of the A549 cells in various groups;5-ethynyl-2'-deoxyuridine(EdU)staining method was used to detect the proliferation abilities of the A549 cells in various groups.After co-culturing A549 cells in various groups with human peripheral blood lymphocytes,flow cytometry was used to detect the percentages of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in various groups;3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)method was used to detect the killing rates of human peripheral blood lymphocytes on the A549 cells in various groups.Results:The diameter of Exo vesicles was 50-150 nm,and the exosome-specific marker proteins cluster of differentiation 63(CD63),cluster of differentiation 9(CD9),tumor susceptibility gene 101(TSG101),and heat shock protein 70(HSP70)were positively expressed,indicating successful exosome extraction.A549 cells efficiently took up PKH67-labeled Exo.The RT-PCR results showed that compared with A549 cells cultured alone,the expression level of lncRNA DUXAP8 in the A549 cells was increased after treatment with Exo derived from A549 cells(P<0.05).compared with control group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there were no significant difference in the expression level of IncRNA DUXAP8 in the cells in Exo+sh-NC group(P>0.05).The colony formation assay results showed that compared with control group,the number of colony formation of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the number of colony formation of the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the number of colony formation of the A549 cells in Exo+sh-NC group(P>0.05).The EdU staining results showed that compared with control group,the EdU-positive rate of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the EdU-positive rate in A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the EDU-positive rate in the cells in Exo+sh-NC group(P>0.05).The flow cytometry results showed that compared with control group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo group was decreased(P<0.05);compared with Exo group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo+sh-DUXAP8 group was increased(P<0.05),while there was no significant difference in the percentage of activated CD8+T lymphaytes in Exo+sh-NC group(P>0.05).The MTT assay results showed that compared with control group,the killing rate of human peripheral blood lymphocytes on the A549 cells in Exo group was decreased(P<0.05);compared with Exo group,the killing rate of human peripheral blood lymphocytes on A549 cells in Exo+sh-DUXAP8 group was increased(P<0.05),while no significant difference was observed in Exo+sh-NC group(P>0.05).Conclusion:The lncRNA DUXAP8 in exosomes derived from the lung cancer A549 cells promotes the proliferation of lung cancer cells and tumor immune escape.
3.Advantages of modified ligation method for spinal cord injury modeling
Daohui LI ; Xiaoshuang XU ; Zhengtao LI ; Xinpeng TIAN ; Hangchuan BI ; Yuan LIU ; Yongwen DAI ; Lingqiang CHEN
Chinese Journal of Tissue Engineering Research 2025;29(2):379-384
BACKGROUND:Currently,different methods of model establishment have been derived from different injury modes of spinal cord injury.Traditional physical injury modeling methods have their own advantages and disadvantages,and there is a lack of more effective and stable animal models of spinal cord injury. OBJECTIVE:To establish a reproducible,controllable,trauma-free,low-mortality,more stable,widely applicable,and short-term postoperative care rat model of spinal cord injury. METHODS:Forty Sprague-Dawley rats with similar body mass and ages were randomly divided into a control group and an improved group,with 20 rats in each group.Animal models of spinal cord injury in the control group were constructed using a clip model method,while the improved group used a modified ligation method based on the compression method to make the spinal cord injury models using suture ligation based on fenestration.Postoperative comparisons were made between the two groups,assessing urination behavior,hematuria,pyuria(infection rate),mortality,scoliosis rate and Basso-Beattie-Bresnahan locomotor rating scale scores at 1,3,5,and 7 days after modeling. RESULTS AND CONCLUSION:Compared with the conventional modeling method,the modified ligation method based on the compression method resulted in faster recovery of urination behavior,lower hematuria rate,lower infection rate,lower mortality rate,lower scoliosis rate,and more concentrated and stable Basso-Beattie-Bresnahan scores(all below 2 points within 1 week).This proves that the modified ligation method based on compression is more suitable for the establishment of spinal cord injury models in rats.
4.Mechanisms of bone marrow mesenchymal stem cells in counteracting D-galactose-induced brain aging
Xiaoxu CHEN ; Xiaoshuang YUAN ; Ting TIAN ; Bingbing LI ; Bo YANG ; Xu YANG ; Tingting TIAN ; Fa CHEN ; Yanju LI ; Dongxin TANG ; Yang LIU ; Feiqing WANG
Acta Laboratorium Animalis Scientia Sinica 2025;33(10):1412-1421
Objective To investigate the effect and potential mechanism of rat mesenchymal stem cells(MSC)on D-galactose-induced brain-tissue aging.Methods A rat brain-aging model was established by injecting D-galactose,and rats in the treatment group received MSC injections via the tail vein.Superoxide dismutase(SOD)activity and malondialdehyde(MDA)levels were assessed in rat brain tissue at the end of the experiment,and pathological changes in brain tissue were observed by hematoxylin-eosin(HE)staining.Expression levels of the inflammatory factors interleukin(IL)-1 and IL-6,the pathway proteins brain-derived neurotrophic factor(BDNF)-tropomyosin receptor kinase B(TrkB),the negative growth regulators p53 and p16,as well as vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)were observed by polymerase chain reaction(PCR)and Western Blot.Results Brain levels of SOD activity were significantly increased and MDA levels were significantly decreased in rats in the modle group compared with the treatment group(P<0.05).The pathological state of the cerebral cortex and hippocampus were improved and the number of neurons and nucleus pulposus ratio in the brain were increased in the treatment group,as shown by HE staining.Expression levels of IL-1,IL-6,p53,and p16 were significantly decreased,while BDNF,TrkB,VEGF,and bFGF were significantly increased in the treatment group compared with the model group,as shown by PCR and Western Blot(P<0.05).Conclusions These result suggest that MSCs potentially mitigate D-galactose-induced cerebral senescence by concurrently modulating the BDNF-TrkB axis to attenuate oxidative/inflammatory damage,while enhancing the secretion of vasculotrophic(VEGF)and neurotrophic(bFGF)factors for neuronal maintenance.
5.The Effect of 1 800 MHz Electromagnetic Radiation on Learning and Cognitive Functions in 3xTg-AD Mice
Xiaoshuang XU ; Guoyu MA ; Runhua HE ; Yongli ZHAO ; Huixin WU ; Yunzhen MU
Journal of Kunming Medical University 2025;46(2):37-43
Objective To explore the effects of 1 800 MHz electromagnetic radiation(EMR)on cognitive function of 3xTg-AD and 57C mice,providing a theoretical basis for the potential impacts of electromagnetic radiation on the human body.Methods A total of 12 3xTg-AD transgenic mice and 12 wild-type C57 mice were selected as research subjects.The one-month-old mice were divided into four groups:RF WT,Control WT,RF AD,and Control AD,with 6 mice in each group.The 1 800 MHz EMR exposure experiments were conducted from 20:00 to 8:00 the next day for a duration of 5 months.After the exposure,a water maze test was conducted to evaluate the effects of EMR on spatial learning and memory abilities of 3xTg AD mice,along with measurements of body weight,brain weight,and calculation of the brain-to-body ratio.Finally,Western Blot technique was used to measure the levels of APP,NR1,and NR2A in hippocampal tissue to analyze effects of 1 800 MHz EMR on the cognitive function of 3xTg AD mice.Results Under 1 800 MHz EMR exposure,there were no statistically significant differences in Morris water maze spatial learning ability among the four groups(P>0.05).However,longer escape time,greater swimming distances,and more crossings of target quadrant were exhibited in the RF AD group compared to the other groups(P<0.05).Western Blot results showed that the APP protein levels in 3xTg AD mice was higher than those in C57 mice(P<0.05).The expression levels of NR1 protein in the WT group was higher than those in the AD group(P<0.05);in the AD group,the RF AD group had higher levels than the Control WT group(P<0.05),and the NR2A protein levels in the Control WT group were higher than in the other groups(P<0.05).Conclusion Prolonged exposure to 1 800 MHz EMR can affect the learning and cognitive function of both 3xTg AD and C57 mice.
6.Association between physical activity and cognitive impairment in older adults aged 65 years and above in longevity areas of China
Hang XU ; Yudong WU ; Chen CHEN ; Xi MENG ; Jiahao CHEN ; Zenghang ZHANG ; Zhuchun ZHONG ; Jingjing YANG ; Xiaoshuang FU ; Sirui CHEN ; Yongqiang CHEN ; Zhipei LI ; Lin YE ; Xiaoming SHI ; Yuebin LYU
Chinese Journal of Epidemiology 2025;46(5):753-760
Objective:To explore the relationships between physical activity and cognitive impairment in older adults aged ≥65 years in longevity areas in China.Methods:A total of 6 081 older adults aged ≥65 years from the Healthy Ageing and Biomarkers Cohort Study in China in 2021 were included in this study. Information about their demographic characteristics, lifestyles, and chronic disease histories were collected, the intensity of physical activity was evaluated by using Physical Activity Scale for the Elderly, and the cognitive function was evaluated by using Mini-Mental State Examination Scale (Chinese version). Multifactorial logistic regression model was used to analyze the associations between different levels and types of physical activity and cognitive impairment in older adults.Results:In the 6 081 older adults, 1 829 (30.1%) had cognitive impairment. After adjusting for confounders, older adults with T2 and T3 levels of physical activity had lower risks for cognitive impairment compared with those with T1 levels of physical activity, with ORs of 0.47 (95% CI: 0.40-0.55) and 0.22 (95% CI: 0.18-0.28). The results of different types of physical activities showed that the ORs in leisure activity T2 and T3 groups were 0.52 (95% CI: 0.44-0.63) and 0.49 (95% CI: 0.41-0.58), and the ORs in housework activity T2 and T3 groups were 0.36 (95% CI: 0.30-0.42) and 0.19 (95% CI: 0.16-0.24). There was no significant association between work-related activity and cognitive impairment. Conclusion:There is a negative association between the intensity level of physical activity and cognitive impairment, and active leisure and household activities might reduce the risk for cognitive impairment.
7.Best evidence summary for prevention and management of high-output ileostomy
Qing ZHANG ; Haiyan HU ; Xin YIN ; Xiaoshuang ZHAO ; Yuting XU ; Tingting HE ; Jianan SUN
Chinese Journal of Practical Nursing 2025;41(29):2301-2307
Objective:To retrieve, appraise and synthesize evidence regarding the prevention and management of high-output ileostomy in post-enterostomy patients, thereby providing a scientific reference for clinical practice.Methods:A computerized search was conducted, systematically exploring national and international databases, as well as the websites of pertinent associations, to gather evidence relevant to the topic. The literature types included clinical decision-making, evidence summaries, guidelines, expert consensus, and systematic reviews. The search timeframe was established from the date of database construction until 23 August 2024. The methodological quality assessment of the literature, along with the extraction and synthesis of the evidence, was performed by two independent researchers utilizing established criteria and adhering to rigorous standards.Results:A total of 13 papers were included in the review, comprising one clinical decision, four guidelines, and eight systematic reviews. Eighteen pieces of best evidence were distilled and summarized across six key areas: diagnosis and risk identification, fluid/diet/nutrition management, medication administration, skin management, monitoring and health education, and follow-up care.Conclusions:The clinical practice rationale for the prevention and management of high-output ileostomy in post-enterostomy patients, when summarized, would offer scientific and effective evidence-based support for practice changes by clinical care staff. The practical application and translation of this evidence should thoroughly consider the nuances of clinical practice in China, as well as individual patient differences, to ensure the effective implementation of the best evidence for maximizing patient benefit.
8.Association between physical activity and cognitive impairment in older adults aged 65 years and above in longevity areas of China
Hang XU ; Yudong WU ; Chen CHEN ; Xi MENG ; Jiahao CHEN ; Zenghang ZHANG ; Zhuchun ZHONG ; Jingjing YANG ; Xiaoshuang FU ; Sirui CHEN ; Yongqiang CHEN ; Zhipei LI ; Lin YE ; Xiaoming SHI ; Yuebin LYU
Chinese Journal of Epidemiology 2025;46(5):753-760
Objective:To explore the relationships between physical activity and cognitive impairment in older adults aged ≥65 years in longevity areas in China.Methods:A total of 6 081 older adults aged ≥65 years from the Healthy Ageing and Biomarkers Cohort Study in China in 2021 were included in this study. Information about their demographic characteristics, lifestyles, and chronic disease histories were collected, the intensity of physical activity was evaluated by using Physical Activity Scale for the Elderly, and the cognitive function was evaluated by using Mini-Mental State Examination Scale (Chinese version). Multifactorial logistic regression model was used to analyze the associations between different levels and types of physical activity and cognitive impairment in older adults.Results:In the 6 081 older adults, 1 829 (30.1%) had cognitive impairment. After adjusting for confounders, older adults with T2 and T3 levels of physical activity had lower risks for cognitive impairment compared with those with T1 levels of physical activity, with ORs of 0.47 (95% CI: 0.40-0.55) and 0.22 (95% CI: 0.18-0.28). The results of different types of physical activities showed that the ORs in leisure activity T2 and T3 groups were 0.52 (95% CI: 0.44-0.63) and 0.49 (95% CI: 0.41-0.58), and the ORs in housework activity T2 and T3 groups were 0.36 (95% CI: 0.30-0.42) and 0.19 (95% CI: 0.16-0.24). There was no significant association between work-related activity and cognitive impairment. Conclusion:There is a negative association between the intensity level of physical activity and cognitive impairment, and active leisure and household activities might reduce the risk for cognitive impairment.
9.Mechanisms of bone marrow mesenchymal stem cells in counteracting D-galactose-induced brain aging
Xiaoxu CHEN ; Xiaoshuang YUAN ; Ting TIAN ; Bingbing LI ; Bo YANG ; Xu YANG ; Tingting TIAN ; Fa CHEN ; Yanju LI ; Dongxin TANG ; Yang LIU ; Feiqing WANG
Acta Laboratorium Animalis Scientia Sinica 2025;33(10):1412-1421
Objective To investigate the effect and potential mechanism of rat mesenchymal stem cells(MSC)on D-galactose-induced brain-tissue aging.Methods A rat brain-aging model was established by injecting D-galactose,and rats in the treatment group received MSC injections via the tail vein.Superoxide dismutase(SOD)activity and malondialdehyde(MDA)levels were assessed in rat brain tissue at the end of the experiment,and pathological changes in brain tissue were observed by hematoxylin-eosin(HE)staining.Expression levels of the inflammatory factors interleukin(IL)-1 and IL-6,the pathway proteins brain-derived neurotrophic factor(BDNF)-tropomyosin receptor kinase B(TrkB),the negative growth regulators p53 and p16,as well as vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)were observed by polymerase chain reaction(PCR)and Western Blot.Results Brain levels of SOD activity were significantly increased and MDA levels were significantly decreased in rats in the modle group compared with the treatment group(P<0.05).The pathological state of the cerebral cortex and hippocampus were improved and the number of neurons and nucleus pulposus ratio in the brain were increased in the treatment group,as shown by HE staining.Expression levels of IL-1,IL-6,p53,and p16 were significantly decreased,while BDNF,TrkB,VEGF,and bFGF were significantly increased in the treatment group compared with the model group,as shown by PCR and Western Blot(P<0.05).Conclusions These result suggest that MSCs potentially mitigate D-galactose-induced cerebral senescence by concurrently modulating the BDNF-TrkB axis to attenuate oxidative/inflammatory damage,while enhancing the secretion of vasculotrophic(VEGF)and neurotrophic(bFGF)factors for neuronal maintenance.
10.Best evidence summary for prevention and management of high-output ileostomy
Qing ZHANG ; Haiyan HU ; Xin YIN ; Xiaoshuang ZHAO ; Yuting XU ; Tingting HE ; Jianan SUN
Chinese Journal of Practical Nursing 2025;41(29):2301-2307
Objective:To retrieve, appraise and synthesize evidence regarding the prevention and management of high-output ileostomy in post-enterostomy patients, thereby providing a scientific reference for clinical practice.Methods:A computerized search was conducted, systematically exploring national and international databases, as well as the websites of pertinent associations, to gather evidence relevant to the topic. The literature types included clinical decision-making, evidence summaries, guidelines, expert consensus, and systematic reviews. The search timeframe was established from the date of database construction until 23 August 2024. The methodological quality assessment of the literature, along with the extraction and synthesis of the evidence, was performed by two independent researchers utilizing established criteria and adhering to rigorous standards.Results:A total of 13 papers were included in the review, comprising one clinical decision, four guidelines, and eight systematic reviews. Eighteen pieces of best evidence were distilled and summarized across six key areas: diagnosis and risk identification, fluid/diet/nutrition management, medication administration, skin management, monitoring and health education, and follow-up care.Conclusions:The clinical practice rationale for the prevention and management of high-output ileostomy in post-enterostomy patients, when summarized, would offer scientific and effective evidence-based support for practice changes by clinical care staff. The practical application and translation of this evidence should thoroughly consider the nuances of clinical practice in China, as well as individual patient differences, to ensure the effective implementation of the best evidence for maximizing patient benefit.


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