1.A study on the association between insulin resistance and genome-wide DNA methylation based on Shanghai monozygotic twins
Jingyuan FENG ; Rongfei ZHOU ; Hongwei LIU ; Zihan HU ; Fei WU ; Huiting WANG ; Junhong YUE ; Zhenni ZHU ; Fan WU
Chinese Journal of Epidemiology 2024;45(7):932-940
Objective:To explore the association between insulin resistance (IR) and genome-wide DNA methylation based on Shanghai twin study.Methods:Monozygotic twins (MZ) from Shanghai were recruited during 2012-2013, 2017-2018, and 2022-2023. Data were collected by questionnaire survey, physical examination and laboratory tests. Genome-wide DNA methylation was quantified. Generalized linear mixed effect model was applied to analyze the association between methylation level at each site and homeostatic model assessment 2-insulin resistance (HOMA2-IR). Non-paired and paired designs were used to assess the association between DNA methylation and phenotype of IR. Cluster analysis was conducted to identify the clusters of top significant sites. Generalized linear regression was performed to examine the differential methylation patterns from clusters.Results:A total of 100 MZ pairs were included in this study. Hypermethylated cg10535199-2q23.1 ( β=0.74%, P=1.51×10 -7, OR=1.06, 95% CI: 1.03-1.09) and ch.17.49619327- SPOP ( β=0.23%, P=7.54×10 -7, OR=1.17, 95% CI: 1.08-1.28) were identified with suggestive significance. After correcting for multiple testing, no sites reached genome-wide significance. There was no statistical significance in the paired analysis. Two clusters with hypomethylated ( β=-0.39%, P<0.001) and hypermethylated ( β=0.47%, P<0.001) patterns were observed for HOMA2-IR. Conclusions:IR was significantly associated with DNA methylation, and genetic factors might contribute to the association.
2.Characteristics of allergen component in dust mite-induced allergic rhinitis patients.
Hao CHEN ; Ping JI ; Lin YANG ; Nan HUANG ; Qing JIANG ; Yaqi YANG ; Rongfei ZHU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2023;37(6):442-447
Objective:To investigate the characteristics of allergen component in dust mite(DM) -induced allergic rhinitis(AR) patients, and provide reference for the diagnosis and treatment of AR. Methods:DM-induced AR patients with or without allergic asthma(AA) who visited the Allergy Department of Tongji Hospital, Huazhong University of Science and Technology between 2021 and 2022 were enrolled. Patients'age, gender, and visual analog scale(VAS) for symptoms were recorded. sIgE and sIgG4 levels of allergen components such as Der f1, Der f2, Der p1, Der p2, Der p7, Der p10, Der p21, and Der p23 were detected using a protein chip method. The sensitization characteristics of the allergen components in the patients were observed, and the correlation between sIgE, sIgG of each component and VAS as well as the component differences between AR and AR with AA(AR&AA) were evaluated. Results:A total of 87 DM-induced AR patients were enrolled, with 42.5% of them were AR&AA, their VAS scores were significantly higher than those of AR patients(6.38±1.95 vs 5.25±1.85, P=0.009 8). The order of sensitization rates for DM components was as follows: Der p2(82.8%), Der f2(81.6%), Der p1(74.7%), Der f1(70.1%), and Der p23(35.6%). The order of positive rates for sIgG4 was: Der p2(21.8%), Der f2(13.8%), Der p21(8.0%), and Der p7(6.9%). There were no correlation between the sIgE, sIgG4 levels or positive numbers of components and VAS scores, but there were positive correlations between sIgE, sIgG4 concentrations of components. Compared with AR patients, AR&AA patients had higher levels of sIgE for Der p(60.5[7.2-91.1]vs 14.0[4.8-45.1], P=0.02), Der f(49.8[15.7-81.6]vs 21.3[7.0-50.2], P=0.04), Der p1(27.2[0.7-51.5]vs 2.6[0.2-24.9], P=0.02), Der p2(20.0[1.4-60.6]vs 5.5[0.6-19.1], P=0.004), and Der f2(58.9[16.0-89.2]vs 23.4[0.9-56.8], P=0.009), and a higher proportion of AR with AA patients had sIgE levels of Der p1(70.3% vs 48.0%, P=0.038) and Der p23(27.0% vs 14.0%, P=0.039) that were ≥3 grades. Conclusion:Der p1/f1, Der p2/f3, and Der p23 are the major components of DM sensitized AR patients. Multiple component sensitization and sIgE, sIgG4 levels of each component are not correlated with the severity of AR. The sIgE levels of the Der p1/f1, Der p2/f3, and Der p23 components in AR&AA patients are higher than AR.
Animals
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Humans
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Allergens
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Pyridinolcarbamate
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Rhinitis, Allergic/therapy*
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Pyroglyphidae
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Asthma
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Antigens, Dermatophagoides
3.Effects of Shenfu yixin granule on mitochondrial autophagy of cardiomyocytes in rats with heart failure after acute yocardial infarction
Yaxuan CAO ; Rongfei ZHENG ; He WANG ; Yingjie CAO ; Wenjie DONG ; Lin CUI ; Bin LI ; Yushan CHEN ; Mingjun ZHU
China Pharmacy 2022;33(10):1183-1188
OBJECTIVE To study the effects of Shenfu yixin granule on mitochondrial autophagy of cardiomyocytes in rats with heart failure after acute myocardial infarction. METHODS The model of heart failure after acute myocardial infarction was established by ligaturing the anterior descending branch of the left coronary artery in rats. The model rats were divided into model group,Shenfu yixin granule low-dose and high-dose groups (1.76,8.8 g/kg),Fosinopril sodium tablets group (positive control ,4 mg/kg),sham operation group was set up (only threading without ligation at the same position ),with 8 rats in each group. After 4 weeks of drug intervention ,the hemodynamic indexes of rats in each group were measured by physiological recorder. The pathological changes of myocardial tissue were observed in each group. The level of oxidative stress in cardiomyocytes , mitochondrial membrane potential ,protein expression of PTEN-induced putative kinase 1(PINK1),E3 ubiquitin ligase Parkin and ubiquitin binding protein P 62 in myocardial tissue of rats in each group were detected. RESULTS Compared with sham operation group ,the pathological injuries such as myocardial fiber morphology disorder and inflammatory cell infiltration were serious. The left ventricular end systolic pressure (LVESP),maximum rate of rise of left ventricular internal pressure (+dp/dtmax), maximun rate of decrease of left ventricular internal pressure (-dp/dtmax),total antioxidant capacity ,mitochondrial membrane potential,PINK1,Parkin and P 62 protein expression were significantly decreased in model group (P<0.01). The left ventricular end diastolic pressure (LVEDP),the level of reactive oxygen species and the activity of reduced nicotinamide adenine dinucleotide phosphate in left ventricular ischemic cardiomyocytes were significantly increased (P<0.01). Compared with model group ,the pathological injuries of myocardial tissue in intervention groups were alleviated ,and above indexes were improved in varying degrees(P<0.01 or P<0.05). CONCLUSIONS Shenfu y ixin granule can reduce the level of oxidative stress and alleviate heart failure after acute myocardial infarction ,which may be related to the activation of Parkin-dependent pathway to strengthen mitochondrial autophagy and reduce mitochondrial dysfunction.
4.Expression of galectin-13 in allergic diseases involving airway, skin and mucous membranes
Lingling YI ; Rongfei ZHU ; Lin YANG ; Lintao HU ; Shuchen ZHANG
Chinese Journal of Preventive Medicine 2022;56(6):831-837
To detect the expression of galectin-13 in allergic diseases and provide a new way for the diagnosis and treatment of allergic diseases. A retrospective analysis method was used to screen 216 patients with allergic diseases with house dust mites or aspergillus as allergens who visited the Department of Allergy and Department of Respiratory of Tongji Hospital attached Tongji Medical College, Huazhong University of Science and Technology from March 2018 to May 2021. These allergic diseases included allergic asthma, allergic bronchopulmonary aspergillosis, allergic rhinitis, allergic conjunctivitis, atopic dermatitis, allergic urticaria. 25 subjects without underlying diseases were selected as healthy controls. The galectin-13 content in serum in each group were detected, and the Pearson correlation was used to determine the correlation between the galectin-13 content in serum in each group and blood eosinophil count, blood specific IgE, the score scale of allergic disease. The expression of Galectin-13 was increased in allergic asthma group (71.44±39.44) pg/ml, allergic bronchopulmonary aspergillosis group (100.10±47.62) pg/ml, allergic rhinitis group (54.11±24.81) pg/ml and dermatitis group (44.12±19.51) pg/ml. The expression of galectin-13 was not significantly increased in allergic urticaria group (32.75±10.29) pg/ml and the allergic conjunctivitis group (30.55±9.87) pg/ml. The galectin-13 content in serum, was positively correlated with blood eosinophil count( r s=0.54, P<0.001) and house dust mite specific IgE ( r s=0.51, P<0.001) in allergic asthma group, and was positively correlated with blood eosinophil count( r s=0.63, P=0.025) and aspergillus fumigatus specific IgE ( r s=0.58, P=0.046) in allergic bronchopulmonary aspergillosis group. It was positively correlated with blood eosinophil count ( r s=0.52, P=0.000 2) and house dust mite specific IgE ( r s=0.41, P=0.005) in allergic rhinitis group. In allergic conjunctivitis group, the expression of galectin-13 was positively correlated with conjunctivitis symptom score ( r s=0.47, P=0.048). In atopic dermatitis group, the expression of galectin-13 was positively correlated with blood eosinophil count ( r s=0.58, P<0.001) and house dust mite specificity IgE ( r s=0.47, P=0.002). In allergic urticaria group, the expression of galectin-13 was not significantly correlated with blood eosinophil count or house dust mite specific IgE. Galectin-13 may be related to the occurrence and progress of allergic diseases and may be involved in the occurrence of eosinophilic inflammation.
5.Expression of galectin-13 in allergic diseases involving airway, skin and mucous membranes
Lingling YI ; Rongfei ZHU ; Lin YANG ; Lintao HU ; Shuchen ZHANG
Chinese Journal of Preventive Medicine 2022;56(6):831-837
To detect the expression of galectin-13 in allergic diseases and provide a new way for the diagnosis and treatment of allergic diseases. A retrospective analysis method was used to screen 216 patients with allergic diseases with house dust mites or aspergillus as allergens who visited the Department of Allergy and Department of Respiratory of Tongji Hospital attached Tongji Medical College, Huazhong University of Science and Technology from March 2018 to May 2021. These allergic diseases included allergic asthma, allergic bronchopulmonary aspergillosis, allergic rhinitis, allergic conjunctivitis, atopic dermatitis, allergic urticaria. 25 subjects without underlying diseases were selected as healthy controls. The galectin-13 content in serum in each group were detected, and the Pearson correlation was used to determine the correlation between the galectin-13 content in serum in each group and blood eosinophil count, blood specific IgE, the score scale of allergic disease. The expression of Galectin-13 was increased in allergic asthma group (71.44±39.44) pg/ml, allergic bronchopulmonary aspergillosis group (100.10±47.62) pg/ml, allergic rhinitis group (54.11±24.81) pg/ml and dermatitis group (44.12±19.51) pg/ml. The expression of galectin-13 was not significantly increased in allergic urticaria group (32.75±10.29) pg/ml and the allergic conjunctivitis group (30.55±9.87) pg/ml. The galectin-13 content in serum, was positively correlated with blood eosinophil count( r s=0.54, P<0.001) and house dust mite specific IgE ( r s=0.51, P<0.001) in allergic asthma group, and was positively correlated with blood eosinophil count( r s=0.63, P=0.025) and aspergillus fumigatus specific IgE ( r s=0.58, P=0.046) in allergic bronchopulmonary aspergillosis group. It was positively correlated with blood eosinophil count ( r s=0.52, P=0.000 2) and house dust mite specific IgE ( r s=0.41, P=0.005) in allergic rhinitis group. In allergic conjunctivitis group, the expression of galectin-13 was positively correlated with conjunctivitis symptom score ( r s=0.47, P=0.048). In atopic dermatitis group, the expression of galectin-13 was positively correlated with blood eosinophil count ( r s=0.58, P<0.001) and house dust mite specificity IgE ( r s=0.47, P=0.002). In allergic urticaria group, the expression of galectin-13 was not significantly correlated with blood eosinophil count or house dust mite specific IgE. Galectin-13 may be related to the occurrence and progress of allergic diseases and may be involved in the occurrence of eosinophilic inflammation.
6.Airborne pollens in Wuhan urban area and its correlation with hospital visits of allergic rhinitis patients from 2017 to 2018
Wenjing LI ; Nan HUANG ; Shuchen ZHANG ; Dongxia MA ; Yaqi YANG ; Rongfei ZHU
Chinese Journal of Preventive Medicine 2021;55(5):620-626
Objective:To investigate the species and concentrations of airborne pollens in Wuhan urban area and their correlation with the number of visits of allergic rhinitis patients.Methods:Retrospective analysis of pollen dispersal characteristics and the number of patients with allergic rhinitis presenting to Tongji Hospital of Huazhong University of Science and Technology in Wuhan city from October 2017 to September 2018, as well as pollen allergen testing results of patients with allergic rhinitis presenting to the Department of Allergy during the same period. Pollen data was collected by a 1-year air sampling conducted in Wuhan City during the same period using the volumetric method. The samples were examined microscopically to identify airborne pollen species and counted, and the concentrations of various pollens were calculated. Information on patients with allergic rhinitis who came to the hospital during the same period was collected, and the correlation between pollen concentration and the number of patient visits was statistically analyzed using Pearson correlation analysis.Results:A total of 35 types of airborne pollen were collected from October 2017 to September 2018. The dominant pollens in spring were Moraceae (68.46%, 1 042/1 522), Pendula (12.22%, 186/1 522) and Cupressaceae (2.30%, 35/1 522); in summer and autumn, the dominant pollens were Artemisia (3.81%, 58/1 522), Humulus (4.01%, 61/1 522) and Ambrosia (0.59%, 9/1 522). The peak number of visits for allergic rhinitis patients occurred in March-April and July-September, both exceeding 2 200 visits and reaching a maximum of 2 545 visits. There was a very weak correlation between the number of visits and the total pollen concentration ( r=0.17, P=0.001). The average monthly pollen skin prick test positive rate of patients with allergic rhinitis was highest in March-May, exceeding 40% with a maximum of 45.73%, and there was a significant correlation between the positive pollen skin prick test positive rate and the average monthly pollen concentration ( r=0.62, P=0.031). Conclusions:Pollen species and concentrations fluctuated continuously with time in Wuhan urban area, with peak pollen dispersal in spring from March to April and in autumn from August to September. The number of visits to patients with allergic rhinitis and the positive pollen skin prick test positive rate increased accordingly during the peak pollen concentration periods.
7.Airborne pollens in Wuhan urban area and its correlation with hospital visits of allergic rhinitis patients from 2017 to 2018
Wenjing LI ; Nan HUANG ; Shuchen ZHANG ; Dongxia MA ; Yaqi YANG ; Rongfei ZHU
Chinese Journal of Preventive Medicine 2021;55(5):620-626
Objective:To investigate the species and concentrations of airborne pollens in Wuhan urban area and their correlation with the number of visits of allergic rhinitis patients.Methods:Retrospective analysis of pollen dispersal characteristics and the number of patients with allergic rhinitis presenting to Tongji Hospital of Huazhong University of Science and Technology in Wuhan city from October 2017 to September 2018, as well as pollen allergen testing results of patients with allergic rhinitis presenting to the Department of Allergy during the same period. Pollen data was collected by a 1-year air sampling conducted in Wuhan City during the same period using the volumetric method. The samples were examined microscopically to identify airborne pollen species and counted, and the concentrations of various pollens were calculated. Information on patients with allergic rhinitis who came to the hospital during the same period was collected, and the correlation between pollen concentration and the number of patient visits was statistically analyzed using Pearson correlation analysis.Results:A total of 35 types of airborne pollen were collected from October 2017 to September 2018. The dominant pollens in spring were Moraceae (68.46%, 1 042/1 522), Pendula (12.22%, 186/1 522) and Cupressaceae (2.30%, 35/1 522); in summer and autumn, the dominant pollens were Artemisia (3.81%, 58/1 522), Humulus (4.01%, 61/1 522) and Ambrosia (0.59%, 9/1 522). The peak number of visits for allergic rhinitis patients occurred in March-April and July-September, both exceeding 2 200 visits and reaching a maximum of 2 545 visits. There was a very weak correlation between the number of visits and the total pollen concentration ( r=0.17, P=0.001). The average monthly pollen skin prick test positive rate of patients with allergic rhinitis was highest in March-May, exceeding 40% with a maximum of 45.73%, and there was a significant correlation between the positive pollen skin prick test positive rate and the average monthly pollen concentration ( r=0.62, P=0.031). Conclusions:Pollen species and concentrations fluctuated continuously with time in Wuhan urban area, with peak pollen dispersal in spring from March to April and in autumn from August to September. The number of visits to patients with allergic rhinitis and the positive pollen skin prick test positive rate increased accordingly during the peak pollen concentration periods.
8.Construction and characterization of a eukaryotic expression vector of full-length and mature IL-37b
Mengyuan LI ; Rongfei WEI ; Xingjiu YANG ; Ruimin ZHU ; Ran GAO
Chinese Journal of Comparative Medicine 2018;28(2):59-63
Objective To construct a eukaryotic expression vector pEGFP N1/IL-37b of full-length and mature IL-37b,and to detect the expression of both full-length and mature IL-37b in RAW 264.7 cells, a mouse macrophage cell line. Methods To construct the eukaryotic vectors of full-length and mature IL-37b by using plasmid pUBC/IL-37b as a template containing the coding region of IL-37b full-length gene. To detect the expression of IL-37b by western blot and confocal microscopy after transfected the recombinant plasmid into RAW 264.7 cells, and to detect the inhibition of full-length and mature IL-37b on IL-6 production by real-time PCR. Results Eukaryotic vectors pEGFP N1/IL-37b expressed full-length and mature IL-37b after transfection in cells, which inhibited LPS-induced IL-6 production. Conclusions Eukaryotic vectors of full-length and mature IL-37b can be successfully constructed,and lays a foundation for further study of anti-inflammation functions and mechanisms of IL-37b.
9.Establishment of two human tumor cell lines with lentivirus-mediated stably Smurf1 silencing and their effect on cell migration
Rongfei WEI ; Jing GUO ; Mengyuan LI ; Ruimin ZHU ; Xingjiu YANG ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(4):46-51
Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration.Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively.After 7 days,the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening,enrichment and expansion.The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot.Transwell assay was used to assess the effect of Smurf1 silencing on cell migration.Results The stable cell lines with Smurf1 silencing are constructed successfully.Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay.Conclusion Smurf1 promotes cell migration.
10.Effect of Smad3 on cell migration of A549 and HeLa cells
Rongfei WEI ; Mengyuan LI ; Xingjiu YANG ; Ruimin ZHU ; Damo XU ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(1):11-15,21
Objective To investigate the effect of Smad3 on cell migration of A549 and HeLa cells.Methods Primers for pCMV-Myc-Smad3 plasmid construction and siRNA targeting Smad 3 were designed and synthesized .pCMV-Myc-Smad3 plasmid was constructed with molecular cloning techniques .Overexpression of Smad 3 with Myc-tag or silencing of endogenous Smad3, and then scratch assay was used to detect the migration ability of A 549 and HeLa cells in vitro. Results pCMV-Myc-Smad3 plasmid was successfully constructed .Overexpression of Smad 3 significantly up-regulated the migration rate of A549 and HeLa cells.Conversely, in the same cells, silencing of endogenous Smad3 or treatment with Smad3 inhibitor, SIS3, down-regulated the migration rate .Conclusions Smad3 promotes cell migration of A549 and HeLa cells.

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