1.miR-207 targets autophagy-associated protein LAMP2 to regulate the mechanism of macrophage-mycobacterium tuberculosis interaction.
Wenya DU ; Yumei DAI ; Linzhi YUE ; Tao MA ; Lixian WU
Chinese Journal of Cellular and Molecular Immunology 2025;41(2):97-104
Objectives miR-207 has been identified as being expressed in natural killer (NK) cell exosomes that play a role in disease progression; however, to date, there are no studies specifically linking miR-207 to tuberculosis (TB). Methods Bioinformatics methods employed for prediction, followed by a dual luciferase reporter assay to determine whether lysosome-associated membrane protein 2 (LAMP2) is targeted by miR-207. The experiments were divided into four groups using the liposome transfection method (OP-LAMP2 group: co-transfected with miR-207 mimics and LAMP2 overexpression plasmid; EP group: co-transfected with mimics NC and null-loaded plasmid; siLAMP2 group: transfected with siLAMP2; and siLAMP2-NC group: transfected with siLAMP2-NC). TB infection was modeled using H37Ra-infected Ana-1 cells. The impact of LAMP2 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria were assessed by tuberculosis colony-forming unit counting. Flow cytometry was used to assess the total apoptosis rate. Real-time fluorescent quantitative PCR was conducted to determine the relative expression of LAMP2, apoptosis genes, pyroptosis genes, and autophagy genes. Western blot analysis was performed to measure the relative expression of LAMP2 proteins, apoptosis proteins, pyroptosis proteins, and autophagy proteins. Results Dual luciferase reporter assay test showed that there was a targeting relationship between LAMP2 and miR-207. The transfection model was successfully constructed under real-time fluorescent quantitative PCR and Western blot statistical analysis, and microscopic observation. The infection model was successfully established under microscopic observation. Colony forming unit counting revealed that the number of colonies in the OP-LAMP2 group was lower than that in the EP group, while the number of colonies in the siLAMP2 group was higher than that in the siLAMP2-NC group. Flow cytometry assay revealed that the total apoptosis in OP-LAMP2 group was lower than that in EP group, and the total apoptosis in siLAMP2 group was higher than that in siLAMP2-NC group. Real-time fluorescence quantitative PCR and Western blot analysis revealed that the relative expression of apoptosis and pyroptosis-related proteins and genes in the control group was lower in the OP-LAMP2 group compared to the EP group, and higher in the siLAMP2 group compared to the siLAMP2-NC group. Real-time fluorescence quantitative PCR detected that the relative expression of autophagy positively regulated genes Microtubule-associated protein 1 light chain 3(LC3)and Beclin1 in the OP-LAMP2 group was higher in the OP-LAMP2 group compared to the EP group, and lower in the siLAMP2 group compared to the siLAMP2-NC group, while the relative expression of negatively regulated autophagy genes followed the opposite trend to that of autophagy positively regulated genes. The relative expression of autophagy-related proteins was consistent with the trend of autophagy genes. Conclusions miR-207 enhances macrophage apoptosis, cellular pyroptosis and inhibits autophagy, promoting survival of Mycobacterium tuberculosis by targeting the autophagy-related protein LAMP2, thus offering a novel therapeutic direction for tuberculosis.
Lysosomal-Associated Membrane Protein 2/metabolism*
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MicroRNAs/metabolism*
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Mycobacterium tuberculosis/physiology*
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Autophagy/genetics*
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Humans
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Macrophages/metabolism*
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Apoptosis/genetics*
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Tuberculosis/metabolism*
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Cell Line
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Pyroptosis/genetics*
2.Advances in the study of exosomes derived from mesenchymal stem cells in the treatment of pulmonary diseases.
Tao MA ; Linzhi YUE ; Yumei DAI ; Wenya DU ; Lixian WU
Chinese Journal of Cellular and Molecular Immunology 2025;41(3):278-282
Pulmonary diseases, as a prevalent category of respiratory system disorders, have become a significant global public health concern. The increasing incidence of these diseases, caused by environmental pollution and occupational hazards, poses a substantial threat to human health and the overall quality of life. Mesenchymal stem cells (MSCs) are known for their remarkable immunomodulatory, anti-bacterial, and anti-apoptotic capabilities. Exosomes derived from MSCs, carrying a diverse array of proteins, lipids, nucleic acids, and other bio-active molecules, have demonstrated considerable therapeutic potential in treating pulmonary diseases, and have come to the forefront of medical research. This review summarized the therapeutic role of exosomes derived from various sources of mesenchymal stem cells in the context of pulmonary diseases, aiming to provide a robust foundation for their clinical application in diagnosis and treatment.
Exosomes/transplantation*
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Humans
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Mesenchymal Stem Cells/metabolism*
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Lung Diseases/therapy*
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Animals
3.Establishment and evaluation of a Nomogram early prediction model for severe dengue fever
Li LIU ; Hongjun LI ; Lixian CHANG ; Hui CHEN ; Zhihui MA ; Zhijian DONG ; Lingjun SHEN ; Chunyun LIU
Chinese Journal of Endemiology 2025;44(3):179-185
Objective:To analyze the influencing factors of severe dengue fever patients in the early stage, construct a early prediction model for severe dengue fever, and evaluate it.Methods:A retrospective analysis was conducted to collect early clinical data of dengue fever patients admitted to the People's Hospital of Mengla County and the Third People's Hospital of Kunming in Yunnan Province from July to December 2023. The multifactor logistic regression was used to analyze the factors affecting the severe dengue fever patients, and Nomogram prediction model was used for visualization. Receiver operating characteristic (ROC) curve and calibration curve analysis were used to evaluate the model.Results:A total of 534 dengue fever patients were included, including 291 males and 243 females, aged (39.95 ± 15.69) years. Among them, there were 59 cases (11.05%) of severe dengue fever. The results of multifactor logistic regression analysis showed that age ( OR = 1.05, 95% CI: 1.02 - 1.08, P < 0.001), cardiovascular disease ( OR = 5.28, 95% CI: 2.08 - 13.40, P < 0.001), serous effusion ( OR = 4.34, 95% CI: 1.63 - 11.57, P = 0.003), aspartate aminotransferase ( OR = 1.03, 95% CI: 1.02 - 1.04, P < 0.001), lactate dehydrogenase ( OR = 1.00, 95% CI: 1.00 - 1.01, P = 0.001), and fibrinogen ( OR = 0.46, 95% CI: 0.28 - 0.76, P = 0.003) were independent influencing factors in the early stage of severe dengue fever. The area under the ROC curve of the Nomogram prediction model constructed from the above six variables was 0.96 (0.93 - 0.98). The calibration curve analysis results showed that the mean absolute error between the predicted values of the Nomogram prediction model and the actual observed values was 0.014. Conclusions:Age, cardiovascular disease, serous effusion, aspartate aminotransferase, lactate dehydrogenase, and fibrinogen are independent influencing factors in the early stage of severe dengue fever. The Nomogram prediction model established based on these variables has good predictive ability for severe dengue fever.
4.Establishment and evaluation of a Nomogram early prediction model for severe dengue fever
Li LIU ; Hongjun LI ; Lixian CHANG ; Hui CHEN ; Zhihui MA ; Zhijian DONG ; Lingjun SHEN ; Chunyun LIU
Chinese Journal of Endemiology 2025;44(3):179-185
Objective:To analyze the influencing factors of severe dengue fever patients in the early stage, construct a early prediction model for severe dengue fever, and evaluate it.Methods:A retrospective analysis was conducted to collect early clinical data of dengue fever patients admitted to the People's Hospital of Mengla County and the Third People's Hospital of Kunming in Yunnan Province from July to December 2023. The multifactor logistic regression was used to analyze the factors affecting the severe dengue fever patients, and Nomogram prediction model was used for visualization. Receiver operating characteristic (ROC) curve and calibration curve analysis were used to evaluate the model.Results:A total of 534 dengue fever patients were included, including 291 males and 243 females, aged (39.95 ± 15.69) years. Among them, there were 59 cases (11.05%) of severe dengue fever. The results of multifactor logistic regression analysis showed that age ( OR = 1.05, 95% CI: 1.02 - 1.08, P < 0.001), cardiovascular disease ( OR = 5.28, 95% CI: 2.08 - 13.40, P < 0.001), serous effusion ( OR = 4.34, 95% CI: 1.63 - 11.57, P = 0.003), aspartate aminotransferase ( OR = 1.03, 95% CI: 1.02 - 1.04, P < 0.001), lactate dehydrogenase ( OR = 1.00, 95% CI: 1.00 - 1.01, P = 0.001), and fibrinogen ( OR = 0.46, 95% CI: 0.28 - 0.76, P = 0.003) were independent influencing factors in the early stage of severe dengue fever. The area under the ROC curve of the Nomogram prediction model constructed from the above six variables was 0.96 (0.93 - 0.98). The calibration curve analysis results showed that the mean absolute error between the predicted values of the Nomogram prediction model and the actual observed values was 0.014. Conclusions:Age, cardiovascular disease, serous effusion, aspartate aminotransferase, lactate dehydrogenase, and fibrinogen are independent influencing factors in the early stage of severe dengue fever. The Nomogram prediction model established based on these variables has good predictive ability for severe dengue fever.
5.Clinical significance of serum antioxidant markers in cephalofacial herpes zoster
Qianyang ZHOU ; Xinxin MA ; Xingping ZHENG ; Lixian XU ; Ruili ZHANG
International Journal of Laboratory Medicine 2024;45(22):2694-2698
Objective To investigate the expression change of serum antioxidant markers in cranofacial herpes zoster and its relationship with clinical severity and postherpetic neuralgia(PHN).Methods Totally 109 cases of cranofacial herpes zoster(cranofacial group)and 169 cases of non-cranofacial herpes zoster(non-cranofacial grlup)hospitalized in the Department of Dermatology in the Second Affiliated Hospital of Nanjing Medical University and 200 cases of health check-ups without underlying diseases(healthy control group)from January 2022 to December 2023 were selected,and the serum antioxidant markers including uric acid(UA),total bilirubin(TBIL),and albumin(ALB)of the patients in the three groups group were compared.Meanwhile,the relationship between these markers and clinical severity and PHN of cranofacial herpes zoster was assessed.Results The serum levels of UA,TBIL and ALB in the cranofacial group and non-cranofacial were significantly lower than those in the healthy control group(P<0.05).Serum UA,TBIL and ALB levels in cranofacial group were significantly lower than those in non-cranofacial group(P<0.05),and were nega-tively correlated with disease severity(P<0.05).Serum UA,TBIL and ALB levels in cranofacial group were independent influencing factors for the occurrence of PHN(P<0.05),and had predictive value for PHN in cranofacial herpes zoster(P<0.05).Conclusion Reactivation of cranofacial herpes zoster virus,acute nerve injury and PHN may all be related to the low antioxidant status of the body,and the antioxidant biomarkers UA,TBIL and ALB may be protective factors for herpes zoster,but more studies are needed to clarify the un-derlying mechanisms.
6.Research progress on the role of ferroptosis in infectious diseases
Linzhi YUE ; Tao MA ; Yumei DAI ; Wenya DU ; Guofu WANG ; Lixian WU
Chinese Journal of Comparative Medicine 2024;34(7):175-180
Ferroptosis is a newly discovered mode of programmed cell death characterized by the accumulation of intracellular iron-dependent lipid peroxidation.Current research has mainly focused on the role of ferroptosis in the field of cancer,but increasing evidence shows that ferroptosis is also related to the occurrence of infectious diseases.Ferroptosis has accordingly been detected in cases of COVID-19,tuberculosis,and cryptococcal meningitis,as well as other diseases.This article reviews the role of ferroptosis in infectious diseases,to provide new ideas for the prevention and treatment of ferroptosis-related infectious diseases.
7.Mechanistic study of miR-207 regulation of Mycobacterium tuberculosis survival in macrophages
Wenya DU ; Yumei DAI ; Linzhi YUE ; Tao MA ; Lixian WU
Chinese Journal of Comparative Medicine 2024;34(12):41-49
Objective miR-207 is differentially expressed in many diseases.We investigated the mechanism by which miR-207 overexpression regulates the survival of Mycobacterium tuberculosis(H37Ra)in macrophages,to provide a theoretical basis for the targeted therapy of tuberculosis.Methods Macrophages were divided into four groups:blank(Ana-1 cells),control(cells infected with H37Ra),mi(infected with H37Ra and transfected with miRNA-207 mimics),and mi-NC(infected with H37Ra and transfected with NC mimics)groups.A model of tuberculosis infection was established using H37Ra-infected Ana-1 cells,and miRNA-207 and NC mimics were transfected into Ana-1 cells using the liposome transfection method.Tuberculosis colony-forming units were counted to assess the effect of miR-207 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria.The total apoptosis rate was detected by flow cytometry.The relative expression levels of miR-207 and apoptosis,pyroptosis,inflammation,and autophagy genes were measured by quantitative real-time polymerase chain reaction(qPCR).Relative expression levels of apoptosis,pyroptosis,and autophagy proteins were detected by Western blot.Fluorescence microscopy and multifunctional enzyme labeling were used to detect the fluorescence intensity of intracellular reactive oxygen species(ROS)and lactate dehydrogenase(LDH).Results Successful establishment of the infection model was observed under the microscope.qPCR showed that miR-207 expression was lower in the control compared with the blank group(P<0.01),indicating differential expression between these two groups.miR-207 expression was significantly higher in the mi compared with the mi-NC group(P<0.0001),indicating successful establishment of the transfection model.The number of colonies and total apoptosis were both higher in the mi group compared with the mi-NC and control groups(P<0.001).qPCR and Western blot showed that the relative expression levels of apoptotic genes and proteins were higher in the control group than in the blank group(P<0.05),and higher in the mi group than in the mi-NC group(P<0.05).The relative expression levels of inflammatory genes were higher in the control than in the blank group(P<0.001).The relative expression levels of inflammatory genes were higher in the mi group than in the mi-NC group(P<0.05),and the relative expression levels of pyroptosis genes and proteins were higher in the control group compared with the blank group(P<0.01)and higher in the mi group compared with the mi-NC group(P<0.05).The relative expression levels of the autophagy positively-regulated genes LC3 and Beclin1 were higher in the control compared with the blank group(P<0.0001),and lower in the mi than in the mi-NC group(P<0.05),while negatively-regulated autophagy genes showed the opposite trend.Autophagy-related proteins showed similar trends to the autophagy genes.ROS fluorescence intensity was higher in the control compared with the blank group(P<0.05),and higher in the mi compared with the mi-NC group(P<0.001).LDH content was higher in the control than in the blank group(P<0.01),but there was no significant difference between the mi and mi-NC groups(P>0.05).Conclusions miR-207 overexpression promotes apoptosis,cellular pyroptosis,and inflammation,inhibits autophagy,and favors H37Ra survival.These result provide a potential new direction for the treatment of tuberculosis.
8.Mechanistic study of miR-207 regulation of Mycobacterium tuberculosis survival in macrophages
Wenya DU ; Yumei DAI ; Linzhi YUE ; Tao MA ; Lixian WU
Chinese Journal of Comparative Medicine 2024;34(12):41-49
Objective miR-207 is differentially expressed in many diseases.We investigated the mechanism by which miR-207 overexpression regulates the survival of Mycobacterium tuberculosis(H37Ra)in macrophages,to provide a theoretical basis for the targeted therapy of tuberculosis.Methods Macrophages were divided into four groups:blank(Ana-1 cells),control(cells infected with H37Ra),mi(infected with H37Ra and transfected with miRNA-207 mimics),and mi-NC(infected with H37Ra and transfected with NC mimics)groups.A model of tuberculosis infection was established using H37Ra-infected Ana-1 cells,and miRNA-207 and NC mimics were transfected into Ana-1 cells using the liposome transfection method.Tuberculosis colony-forming units were counted to assess the effect of miR-207 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria.The total apoptosis rate was detected by flow cytometry.The relative expression levels of miR-207 and apoptosis,pyroptosis,inflammation,and autophagy genes were measured by quantitative real-time polymerase chain reaction(qPCR).Relative expression levels of apoptosis,pyroptosis,and autophagy proteins were detected by Western blot.Fluorescence microscopy and multifunctional enzyme labeling were used to detect the fluorescence intensity of intracellular reactive oxygen species(ROS)and lactate dehydrogenase(LDH).Results Successful establishment of the infection model was observed under the microscope.qPCR showed that miR-207 expression was lower in the control compared with the blank group(P<0.01),indicating differential expression between these two groups.miR-207 expression was significantly higher in the mi compared with the mi-NC group(P<0.0001),indicating successful establishment of the transfection model.The number of colonies and total apoptosis were both higher in the mi group compared with the mi-NC and control groups(P<0.001).qPCR and Western blot showed that the relative expression levels of apoptotic genes and proteins were higher in the control group than in the blank group(P<0.05),and higher in the mi group than in the mi-NC group(P<0.05).The relative expression levels of inflammatory genes were higher in the control than in the blank group(P<0.001).The relative expression levels of inflammatory genes were higher in the mi group than in the mi-NC group(P<0.05),and the relative expression levels of pyroptosis genes and proteins were higher in the control group compared with the blank group(P<0.01)and higher in the mi group compared with the mi-NC group(P<0.05).The relative expression levels of the autophagy positively-regulated genes LC3 and Beclin1 were higher in the control compared with the blank group(P<0.0001),and lower in the mi than in the mi-NC group(P<0.05),while negatively-regulated autophagy genes showed the opposite trend.Autophagy-related proteins showed similar trends to the autophagy genes.ROS fluorescence intensity was higher in the control compared with the blank group(P<0.05),and higher in the mi compared with the mi-NC group(P<0.001).LDH content was higher in the control than in the blank group(P<0.01),but there was no significant difference between the mi and mi-NC groups(P>0.05).Conclusions miR-207 overexpression promotes apoptosis,cellular pyroptosis,and inflammation,inhibits autophagy,and favors H37Ra survival.These result provide a potential new direction for the treatment of tuberculosis.
9.Causes analysis of blood donor deferral in 20 domestic blood centers
Dongyan ZHAO ; Bing JU ; Hai QI ; Heng ZHANG ; Lixian MA ; Rong GUO ; Ling HOU ; Lin BAI ; Yang ZHANG ; Tao QI ; Yang CHEN ; Wenjie HU ; Xiaojun XU ; Rui CHEN ; Lin WANG ; Tao LI ; Wei LUO ; Ning CHENG ; Honghua LIU ; Junying LI ; Yan QIU
Chinese Journal of Blood Transfusion 2022;35(4):360-364
【Objective】 To investigate the main causes of blood donor deferral in domestic blood center. 【Methods】 The causes of donor deferral were classified into 12 categories as previous medical history, drug use, alcohol consumption, menstrual period, underweight, abnormal blood pressure, abnormal body temperature, abnormal hemoglobin (Hb), lipemic blood, positive hepatitis B surface antigen (HBsAg), elevated alanine aminotransferase (ALT) and others according to the comparison indicators of Asia-Pacific Blood Network (APBN) and the national standard Blood Donor Health Examination Requirements. The relevant data of the top 3 causes of donor deferral, voluntarily reported by the members of Practice Comparison Working Group of China’s Mainland Blood Collection and Supply Institutions from 2014 to 2019, were collected and a histogram was generated. 【Results】 The median donor deferral rate of 20 domestic blood centers from 2014 to 2019 was 12.14%, with the lowest at 0.18% and highest at 32.32%, respectively. The top three causes for donor deferral were elevated ALT, abnormal Hb and abnormal blood pressure in year 2014, 2015, 2018 and 2019; elevated ALT, lipemic blood and abnormal blood pressure in 2016; elevated ALT, abnormal Hb, and lipemic blood in 2017. 【Conclusion】 The main causes of donor deferral were elevated ALT, abnormal Hb, abnormal blood pressure and lipemic blood.
10.Recent developments in multimodality fluorescence imaging probes.
Jianhong ZHAO ; Junwei CHEN ; Shengnan MA ; Qianqian LIU ; Lixian HUANG ; Xiani CHEN ; Kaiyan LOU ; Wei WANG
Acta Pharmaceutica Sinica B 2018;8(3):320-338
Multimodality optical imaging probes have emerged as powerful tools that improve detection sensitivity and accuracy, important in disease diagnosis and treatment. In this review, we focus on recent developments of optical fluorescence imaging (OFI) probe integration with other imaging modalities such as X-ray computed tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET), single-photon emission computed tomography (SPECT), and photoacoustic imaging (PAI). The imaging technologies are briefly described in order to introduce the strengths and limitations of each techniques and the need for further multimodality optical imaging probe development. The emphasis of this account is placed on how design strategies are currently implemented to afford physicochemically and biologically compatible multimodality optical fluorescence imaging probes. We also present studies that overcame intrinsic disadvantages of each imaging technique by multimodality approach with improved detection sensitivity and accuracy.

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