1.Dynamic characteristics of intracellular complement components mRNA in different mouse macrophage ANA-1 polarization
Mengjiao YUAN ; Zhengling SHANG ; Yaping MA ; Jiahong WU
Chinese Journal of Immunology 2016;32(6):782-786
Objective:To reveal the dynamic characteristics of the intracellular complement mRNA from mouse macrophage ANA-1 treated with LPS or IL-4. Methods:The polarization models of macrophage ANA-1 were established by treating with LPS(1μg/ml) and IL-4(20 ng/ml),respectively. After treating at 3,8,12 and 24 h,the total RNA were abstracted by Trizol lysis methods . The macrophage polarization were estimated by the expression of IL-1β, CCL2 and Arg-1 mRNA detected by Real-time fluorescent quantitative PCR. The intracellular complement C1q, C3, CfB and CRIg mRNA were quantitatively analyzed. Results: The mouse macrophage ANA-1 cells treated with LPS was polarized to M1 since the levels of IL-1β and CCL2 mRNA were up-regulated significantly,in which their 2-△△Ct value were up to 297. 0±31. 0 and 19. 9±3. 3 respectively at 12 h. On the other hand,the ANA-1 cells treated with IL-4 was polarized to M2 because the level of Arg-1 mRNA was obviously higher( the 2-△△Ct value of Arg-1 mRNA was up to 27.3±9.1 at 24 h)(P<0.05).The intracellular complement C1q,C3,CfB and CRIg mRNAs all were up-regulated in different polarized macrophages. The intracellular C1q and C3 mRNA in polarized M2 were significantly higher,in which the peak value of C1q and C3 were to 94. 9±12. 9 and 11. 3±2. 4 at 12 h,respectively(P<0. 05). Reversely,the CfB mRNA in polarized M1 increased obviously,in which its 2-△△Ct was to 61. 4±6. 2 at 12 h. In addition,the CRIg mRNA in both groups was only up-regulated at 24 h,in which the 2-△△Ct value was 6. 5±1. 8 in M1 and 10. 8±3. 2 in M2(P<0. 05). Conclusion: The macrophage ANA-1 cell polarization models were successfully established by treated with LPS or IL-4. The intracellular complement C1q,C3 and CRIg mRNA in polarized M2 were transcripted more than in M1. But the intracellular CfB mRNA in polarized M1 was up-regulated significantly. These results suggested that the dynamic characteristic of complement components in different polarized macrophage would be correlated with its fun-tions.
2.Diagnostic value of enhanced CT for necrotizing pneumonia in children
Shuaishuai LIU ; Jing MA ; Zhongxiao ZHANG ; Xiuli YAN ; Mengjiao ZHAO ; Na LIU ; Chen MENG
Chinese Journal of Applied Clinical Pediatrics 2021;36(4):267-270
Objective:To study the diagnostic value of non-enhanced lesions on enhanced CT in lung consolidation for necrotizing pneumonia in children.Methods:A total of 101 cases of necrotizing pneumonia with air sacs on CT scan who were hospitalized in the Department of Respiratory Intervention, Qilu Children′s Hospital of Shandong University from August 2016 to September 2018 were enrolled in this study(group with air lucency). Besides, another 75 cases of lobar pneumonia with non-enhanced lesions in lung consolidation but without air sacs on enhanced CT were also included from the same hospital over the same period(group without air lucency). Clinical data of these patients were retrospectively collected and statistically analyzed.Results:The white blood cell count was (12.5±5.5)×10 9/L in group with air sacs and (10.8±4.1)×10 9/L in group without air sacs, and the difference was statistically significant( t=-2.161, P=0.032). There was no statistical difference between the group with and without air sacs in age, gender distribution, the course prior to admission, duration of fever after admission, length of hospital stay, medical expense, the neutrophils percentage in peripheral blood, C-reactive protein, the erythrocyte sedimentation rate, serum procalcitonin, serum D-Dimer, serum lactate dehydrogenase, serum albumin, bronchoscopy times, the bronchial mucosal erosion ratio, the mucus plug score, the lavage purulent lavage ratio, and the ratio of luminal stricture or atresia in late bronchoscopy(all P>0.05). Conclusions:The clinical course of patients with non-enhanced lesions in lung consolidation but without air sacs is almost identical to that of patients with air sacs on CT scan.The presence of non-enhanced lesions in lung consolidation can be used as diagnostic basis of necrotizing pneumonia in children.
3.Research progress on the mechanism of 5-hydroxytryptamine in sepsis
Zhuanhuan DU ; Li MA ; Lingling ZHEN ; Mengjiao SUN ; Xiaorong DONG ; Hong AN
Chinese Critical Care Medicine 2019;31(5):662-664
Sepsis has the characteristics of high morbidity and high mortality, which is the main cause of death in critically ill patients. Peripheral 5-hydroxytryptamine (5-HT) is mainly synthesized and secreted by enterochromaffin cells and plays a role in the development and progression of inflammation. This article focuses on 5-HT, 5-HT transporter (SERT) and selective serotonin reuptake inhibitors (SSRIs) in immune response, inflammatory factor release, oxidative stress, intestinal bacterial translocation and microcirculation of sepsis. The role of the review is to find new ideas for the clinical treatment of sepsis.
4.Study on Myocardial Lymphangiogenesis Disorder in Dilated Cardiomyopathy Mice Induced by Doxorubicin and the Protective Mechanism of Kuoxin Decoction
Yidan DONG ; Mengjiao MA ; Longping PENG ; Maolin ZHOU ; Qianqian LIANG ; Qiong WU ; Yanwen WANG ; Youhua WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(10):3293-3303
Objective To observe the dynamic changes of cardiac lymphangiogenesis in Doxorubicin(DOX)-induced dilated cardiomyopathy(DCM)model mice,and to study the the protective mechanism of Kuoxin Decoction.Methods The DCM mouse model was established by intraperitoneal injection of DOX,and the dynamic observation was performed every week.On this basis,60 C57BL/6 mice were randomly divided into 6 groups(n=10):control group,Model group,L-KXD,M-KXD and H-KXD groups and Captopril group.After successful modeling,the KXD and the positive control drug Captopril were administered continuously for 28 days.Echocardiography was used to detect cardiac function in mice,HE staining and Masson staining were used to observe pathological and morphological changes of the heart,Whole-mount immunofluorescent staining was used to detect the expression of LYVE-1 and Podoplanin in epicardial lymphatic vessels,Western blot was used to detect the expression of VEGFR-3 protein,and qPCR was used to detect the expression of VEGFR-3 mRNA.Results DCM mice induced by DOX showed significant cardiac function decline from the third week(DOX:15 mg·kg-1,P<0.05),and significant ventricular remodeling at the fifth week(DOX:15 mg·kg-1,P<0.01);The lymphatic vessel area of the mouse heart decreased significantly from the fourth week(DOX:20 mg·kg-1,P<0.0001),and the expression of VEGFR-3 decreased significantly from the third week(DOX:15 mg·kg-1,P<0.01).Conclusion KXD can improve ventricular remodeling and cardiac function in DOX-induced DCM mice,promote cardiac lymphangiogenesis,and upregulate the expression of VEGFR-3 at protein and mRNA levels,with a better effect than captopril.DOX-induced cardiac lymphangiogenesis in DCM mice leads to severe myocardial fibrosis and weakened cardiac function,which gradually worsens with the accumulation of modeling time and dose.KXD can promote cardiac lymphangiogenesis and improve cardiac function in DOX-induced DCM mice.The mechanism may be related to the up-regulation of VEGFR-3 expression.
5.In vitro study of dendritic cell derived exosomes transporting hepatitis D antigen induced specific cytotoxic T lymphocyte response
Ting YAO ; Mengjiao LYU ; Jinmei CHEN ; Yi ZHANG ; Siyuan MA ; Yongsheng YU ; Guoqing ZANG ; Xiaohua CHEN
Chinese Journal of Infectious Diseases 2022;40(4):234-240
Objective:To explore the effect and mechanism of dendritic cell derived exosomes (Dexs) loading ubiquitinated (Ub) hepatitis D antigen (HDAg) on activating specific cytotoxic T lymphocytes (CTL).Methods:Ub-S-HDAg-Dexs were co-cultured with dendritic cells (DC) which were from the femora of C57BL/6 mice for 48 h, then flow cytometry was used to detect the maturity of DC (CD86, CD80 and major histocompatibility complex (MHC) Ⅱ). The spleen-derived T lymphocytes from C57BL/6 mice were added in vitro to activate DC and co-cultivated for 72 h. The T cells were divided into Ub-S-HDAg-Dexs group (add 50 μg/mL Ub-S-HDAg-Dexs), Blank-Dexs group (add 50 μg/mL DC derived exosomes without plasmid transfection), Con-Dexs group (add 50 μg/mL DC derived exosomes transfected by cantrol plasmid), PBS group (add 50 μL/mL phosphate buffered saline), and Ub-S-HDAg-Dexs+ AG490 group (add 50 μg/mL Ub-S-HDAg-Dexs, DC and T lymphocytes stimulated by exosomes, and 50 μmol/L AG490 was also added to the cell mix). Flow cytometry was used to detect CD8 + T cells secreting interferon-gamma, non-radioactive lactate dehydrogenase release test to detect the killing activity of specific CTL. Real-time quantitative polymerase chain reaction (PCR) and Western blotting were used to detect the mRNA and protein expressions of JAK kinase (JAK) 2, GATA-binding protein 3 (GATA3), T-bet, signal transduction and activator of transcription (STAT) 1 and STAT4. Independent sample t test were used for statistical analysis. Results:The positive rates of the surface molecules CD80, CD86, MHCⅡof DC stimulated by Ub-S-HDAg-Dexs were 83.850%±0.219%、68.910%±0.134%、84.320%±0.445%, respectively.In the Ub-S-HDAg-Dexs group, the rate of CD8 + T cells secreting interferon-gamma was 6.420%±0.028%, which was higher than those of other groups, including PBS group, Blank-Dexs group, Con-Dexs group and Ub-S-HDAg-Dexs+ AG490 group ( t=90.78, 30.32, 63.06 and 85.42, respectively, all P<0.001). The cytotoxicity of T cells in the Ub-S-HDAg-Dexs group was 82.4%±3.9%, which was higher than those of other groups ( t=17.28, 9.74, 3.95 and 15.89, respectively, all P<0.050). The relative mRNA expressions of JAK2, T-bet, STAT1, STAT4 in Ub-S-HDAg-Dexs group were higher than those in other groups, including Con-Dexs group ( t=10.74, 32.34, 13.00 and 16.28, respectively, all P<0.001), Blank-Dexs group ( t=15.05, 21.51, 6.46 and 13.12, respectively, all P<0.050), PBS group ( t=21.83, 41.42, 7.30 and 17.50, respectively, all P<0.050), Ub-S-HDAg-Dexs+ AG490 group ( t=35.75, 20.69, 17.02 and 17.07, respectively, all P<0.001), and the differences were all statistically significant. The protein expressions of T-bet, STAT1, STAT4 in Ub-S-HDAg-Dexs group increased compared with those in PBS group ( t=346.70, 57.54 and 55.81, respectively, all P<0.001), with statistical significance. In the presence of AG490, the protein expressions of T-bet, STAT1 and STAT4 decreased compared with those in Ub-S-HDAg-Dexs group, and the differences were statistically significant ( t=355.40, 52.79 and 126.10, respectively, all P<0.001). Conclusions:Ubiquitinated HDAg transported by exosomes could effectively promote DC maturation, induce T lymphocyte differentiation, and generate specific CTL responses, which provides a new idea for the treatment of hepatitis D.
6.The effects of combining virtual reality training with aromatherapy on Alzheimer′s disease patients in a nursing home
Zhicheng SUN ; Tong WANG ; Xiaomei GU ; Jinlin MA ; Na ZHANG ; Mengjiao YANG ; Yun WANG ; Yajun DONG
Chinese Journal of Physical Medicine and Rehabilitation 2023;45(3):226-231
Objective:To explore any effects of combining virtual reality training with aromatherapy in caring for Alzheimer′s disease patients in a nursing home.Methods:Fifty nursing home residents with Alzheimer′s were divided at random into an observation group and a control group, each of 25. Both groups received routine rehabilitation, while the observation group was additionally given 45 minutes of virtual reality training combined with aromatherapy, 3 times a week for 6 months. Both groups′ cognition was then evaluated using the MMSE and an Alzheimer′s cognition assessment scale (ADAS-cog). Psycho-behavioral symptoms were quantified using the Alzheimer′s disease pathological behavior scale (BEHAVE-AD). Motor functioning was quantified using the timed up and go test (TUGT), the 30-second sit-to-stand test (30sCST), the 30-second arm curl test (30sACT) and the sit-and-reach test (CSRT). Ability in the activities of daily living (ADL) and life quality were quantified using the activity of daily living scale and of the quality of life scale for Alzheimer′s disease (QOL-AD) before and after the intervention.Results:After the intervention the average MMSE, ADAS-cog, BEHAVE-AD and ADL scores of both groups had improved significantly, with the average improvement in the observation group significantly greater than that in the control group. The TUGT, 30sCST, 30sACT and CSRT results of both groups were also significantly better, with those of the observation group again significantly superior, on average, to the control group′s results. The average QOL-AD score in the observation was significantly improved after the intervention, and was then significantly better than the control group′s average.Conclusions:Virtual reality training combined with aromatherapy can significantly improve the cognition, psycho-behavioral symptoms, activity in daily living, motor functioning and life quality of Alzheimer′s patients in a nursing home. It is worthy of promotion and application in nursing homes.
7.The clinical value of procalcitonin in predicting the death of patients with acute biliary pancreatitis
Zaiqian CHE ; Bing ZHAO ; Yihui WANG ; Huihui ZHU ; Yuming WANG ; Xing QI ; Mengjiao LI ; Lili XU ; Yuhua ZHOU ; Ying CHEN ; Li MA ; Yi XIA ; Zhiwei XU ; Erzhen CHEN ; Enqiang MAO
Chinese Journal of Pancreatology 2020;20(2):114-119
Objective:To explore the clinical value of procalcitonin (PCT) in predicting mortality of patients with acute biliary pancreatitis (ABP).Methods:The clinical data of 196 ABP patients admitted in the emergency department of Ruijin Hospital Affiliated to Shanghai Jiaotong University Medical College from January 2013 to June 2017 were analyzed retrospectively. The enrolled patients were divided into survival group ( n=176) and death group ( n=20) according to clinical outcome, and their clinical characteristics, laboratory results(including WBC, CRP, PCT), APACHEⅡ score, BISAP score, modified Marshall score, SOFA score and CTSI at admission were compared between two groups. The ROC curve and AUC were used to evaluate the effectiveness of PCT and multiple scoring systems in predicting mortality in ABP patients, and the Delong test was used to compare the predictive efficacy of various methods at 1-2 d, 3-4 d, and 5-7 d days after onset. Results:The PCT level, APACHEⅡ score, BISAP score, modified Marshall score, SOFA score, and CTSI of patients in the death group were significantly higher than those in the survival group [6.98(3.12, 13.64) μg/L vs 0.55(0.17, 1.74) μg/L, 12.00(6.00, 18.75) vs 6.00(3.00, 9.00), 3.20±1.47 vs 1.59±1.05, 2.85±0.37 vs 1.96±0.64, 5.50(4.00, 9.50) vs 2.00(1.00, 4.25), 5.05±2.33 vs 3.39±1.74], and all the differences were statistically significant (all P values <0.05). The AUC of PCT for predicting death was 0.881 (95% CI 0.820-0.938)and the cut-off value was 2.44. The predictive value of PCT was similar to that of the modified Marshall score, BISAP score and SOFA score, but higher than that of APACHEⅡ score and CTSI (all P values <0.05). The predictive AUC of PCT at 3-4 days after onset was higher than that of modified Marshall score, BISAP score and SOFA score, and were significantly higher than those at 1-2 days after onset. Conclusions:PCT can be used to predict the mortality of ABP within 7 days of onset. The predictive value of PCT was comparable to the modified Marshall score, BISAP score and SOFA score, and the best predictive time was 3-4 days after onset.
8.Risk model of breast cancer prognosis based on the expression profile of long non-coding RNA
Jinsong WANG ; Chunxiao LI ; Ting WANG ; Jingyao ZHANG ; Yantong ZHOU ; Fangzhou SUN ; Mengjiao CHANG ; Fei MA ; Haijuan WANG ; Haili QIAN
Clinical Medicine of China 2020;36(3):217-222
Objective:To construct a prediction model for the prognosis of breast cancer patients with long non-coding RNA expression characteristics.Methods:To construct a long non-coding RNA(LncRNA) model for predicting the prognosis of breast cancer patients.Methods Analyzing LncRNA expression profiles and clinical characteristics of 1 081 breast cancer patients in the cancer genome atlas (TCGA) database.Performing differential expression analysis and univariate analysis on 112 paired breast cancer and normal breast tissues′ transcriptome sequencing data in the TCGA database, and screened for differentially expressed (DELncRNAs) that significantly correlated with the prognosis of BRCA (To reduce batch effects, sequencing data has been normalized using the DESeq function). One thousand eighty-one breast cancer patients were randomly divided into two groups: training set (541) and validation set (540). Performing Cox proportional hazard regression using DELncRNAs and establishing a multi-LncRNA prognosis model in the training set, followed by proportional hazards assumption test(PH assumption test). Patients were divided into high-risk and low-risk groups based on calculated risk score.Kaplan-Meier method was used for survival analysis, and 540 patients′ data were used for validation.To evaluate the prognostic value of the model in patients with squamous cell carcinoma of the lung and hepatocarcinoma in TCGA database.Gene Set Enrichment Analysis (GSEA) was used to analyze the specific mechanism of lncrna affecting the survival of patients.Results:There were 2815 differentially expressed genes screened by transcriptome sequencing, 91 of which were significantly related to the prognosis of breast cancer patients ( P<0.05). Based on the Cox regression analysis of 91 delncrna expression data from 541 breast cancer patients in training set, a Cox proportional risk regression model was constructed based on 5 LncRNA (training set AUC=0.746, validation set AUC=0.650): AC004551.1, MTOR-AS1, KCNAB1-AS2, FAM230G and LINC01283, and PH assumption test( P=0.388). K-M survival analysis showed that the survival time of high-risk group was significantly worse than that of low-risk group (median survival time: 7.049 and 12.21 years, HR 0.367, 95% CI0.228-0.597, P<0.001), and the survival time of high-risk group was significantly shorter than that of low-risk group (median survival time: 7.57 and 10.85 years, HR 0.412, 95% CI0.214-0.793, P<0.001). Similar prediction results were also obtained in other cancer species of TCGA: lung squamous cell carcinoma ( HR 0.604, 95% CI0.383-0.951, P=0.007) and liver cell carcinoma ( HR 0.551, 95% CI0.307-0.987, P=0.011). GSEA results suggested that the expression patterns of the above five LncRNA were related to the cell cycle regulation of tumor cells. Conclusion:The prognostic model constructed based on expression profile of AC004551.1, MTOR-AS1, KCNAB1-AS2, FAM230G and LINC01283 can be used to predict the prognosis of breast cancer patients, which is helpful to further guide clinical treatment.