1.Hydrogen therapy promotes macrophage polarization to the M2 subtype by inhibiting the NF-κB signaling pathway
Xue GAO ; Shiying NIU ; Guohua SONG ; Lulu LI ; Xiaoyue ZHANG ; Wentao PAN ; Xuetao CAO ; Xinhui ZHANG ; Meili SUN ; Guoli ZHAO ; Yueying ZHANG
Chinese Journal of Radiological Health 2024;33(1):33-39
Objective To investigate the role of hydrogen therapy in reducing radiation-induced lung injury and the specific mechanism. Methods Forty C57BL/6 mice were randomly divided into four groups: normal control group, model group, hydrogen therapy group I, and hydrogen therapy group II. A mouse model of radiation-induced lung injury was established. The pathological changes in the lung tissue of the mice were examined with HE staining. Immunofluorescence staining was used to detect the expression of surface markers of M1 and M2 macrophages to observe macrophage polarization. The expression of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), and IL-10 in the lung tissue was measured by immunohistochemistry. The expression of nuclear factor-kappa B (NF-κB) p65 and phosphorylated NF-κB (P-NF-κB) p65 was measured by Western blot. Results HE staining showed that compared with the control group, the model group exhibited alveolar septal swelling and thickening, vascular dilatation and congestion, and inflammatory cell infiltration in the lung tissue; the hydrogen groups had significantly reduced pathological damage and inflammatory response than the model group, with more improvements in hydrogen group II than in hydrogen group I. Immunohistochemical results showed that compared with those in the control group, the levels of the inflammatory cytokines IL-6 and TNF-α were significantly increased in the model group; the hydrogen groups showed significantly decreased IL-6 and TNF-α levels and a significantly increased level of the anti-inflammatory factor IL-10 than the model group, which were more marked in hydrogen group II than in hydrogen group I. Immunofluorescence results showed that compared with the control group, the expression of the surface marker of M1 macrophages in the model group was significantly upregulated; the hydrogen groups showed significantly downregulated M1 marker and significantly upregulated M2 marker, and hydrogen group II showed significantly increased M2 marker compared with hydrogen group I. Western blot results showed that compared with that in the control group, the ratio of P-NF-κB p65/NF-κB p65 in the model group was significantly increased; the P-NF-κB p65/NF-κB p65 ratio was significantly reduced in the hydrogen groups than in the model group, and was significantly lower in hydrogen group II than in hydrogen group I. Conclusion Hydrogen inhalation therapy may reduce the inflammatory response of radiation-induced lung injury by inhibiting the NF-κB signaling pathway to promote the polarization of the macrophage M1 subtype to the M2 subtype.
2.Generation of skin-derived iPSCs from an Osteogenesis imperfecta patient carrying WNT1c.677C>T mutation
Songjie DU ; Xin GUAN ; Meili ZHANG ; Xiuli ZHAO ; Yue HUANG
Chinese Journal of Medical Genetics 2024;41(1):38-41
Objective:To obtain skin-derived induced pluripotent stem cells (iPSCs) from an Osteogenesis imperfecta (OI) patient carrying WNT1c.677C>T mutation in order to provide a new cell model for investigating the underlying molecular mechanism and stem cell therapy for OI. Methods:The pathogenic variant of the patient was identified by Sanger sequencing. With informed consent from the patient, skin tissue was biopsied, and primary skin fibroblasts were cultured. Skin fibroblasts were induced into iPSCs using Sendai virus-mediated non-genomic integration reprogramming method. The iPSC cell lines were characterized for pluripotency, differentiation capacity, and karyotyping assay.Results:The patient was found to carry homozygous missense c. 677C>T (p.Ser226Leu) mutation of the WNT1 gene. The established iPSC lines possessed self-renewal and capacity for in vitro differentiation. It also has a diploid karyotype (46, XX). Conclusion:A patient-specific WNT1 gene mutation ( WNT1c.677C>T) iPSC line was established, which can provide a cell model for the study of OI caused by the mutation.
3.Dermatophagoides farinae induces conjunctival epithelial cell damage to promote neutrophil migration and neutrophil extracellular traps formation
Meili WU ; Ru YAN ; Wenjun ZHAO
Chinese Journal of Schistosomiasis Control 2023;35(3):271-278
Objective To investigate the mechanisms underlying allergic conjunctivitis caused by conjunctival epithelial cell damage, neutrophil migration and neutrophil extracellular traps (NETs) formation induced by crude extracts of Dermatophagoides farinae mite (CDM). Methods Human conjunctival epithelial cells were stimulated with 500, 1 000, 2 000, 4 000 ng/mL, and the expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-8 were detected using quantitative real-time PCR (qPCR) assay and enzyme-linked immunosorbent assay (ELISA). The culture supernatant of human conjunctival epithelial cells was collected and co-cultured with neutrophils. Neutrophil migration was measured using Transwell migration assay, and the expression of NETs markers myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) was quantified using immunofluorescence staining. Neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and then NETs were collected for treatment of human conjunctival epithelial cells. Cell apoptosis was detected using flow cytometry, and the levels of IL-6, TNF-α, IFN-γ and IL-8 were measured in the cell culture supernatant using ELISA. Results Treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL up-regulated IL-6, TNF-α, IFN-γ and IL-8 expression in human conjunctival epithelial cells. Following treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL, the culture supernatant of human conjunctival epithelial cells promoted neutrophil migration and induced increases in the staining intensity of MPO and CitH3. In addition, increased NETs triggered the apoptosis of human conjunctival epithelial cells and IL-6, TNF-α, IFN-γ and IL-8 secretion in the culture supernatant of human conjunctival epithelial cells. Conclusions CDM induces human conjunctival epithelial cell damages, thereby promoting neutrophil migration and NETs formation, while the release of NETs further aggravates human conjunctival epithelial cell damages.
4.Dermatophagoides farinae induces conjunctival epithelial cell damage to promote neutrophil migration and neutrophil extracellular traps formation.
Meili WU ; Ru YAN ; Wenjun ZHAO
Chinese Journal of Schistosomiasis Control 2023;35(3):271-278
OBJECTIVE:
To investigate the mechanisms underlying allergic conjunctivitis caused by conjunctival epithelial cell damage, neutrophil migration and neutrophil extracellular traps (NETs) formation induced by crude extracts of Dermatophagoides farinae mite (CDM).
METHODS:
Human conjunctival epithelial cells were stimulated with 500, 1 000, 2 000, 4 000 ng/mL, and the expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-8 were detected using quantitative real-time PCR (qPCR) assay and enzyme-linked immunosorbent assay (ELISA). The culture supernatant of human conjunctival epithelial cells was collected and co-cultured with neutrophils. Neutrophil migration was measured using Transwell migration assay, and the expression of NETs markers myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) was quantified using immunofluorescence staining. Neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and then NETs were collected for treatment of human conjunctival epithelial cells. Cell apoptosis was detected using flow cytometry, and the levels of IL-6, TNF-α, IFN-γ and IL-8 were measured in the cell culture supernatant using ELISA.
RESULTS:
Treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL up-regulated IL-6, TNF-α, IFN-γ and IL-8 expression in human conjunctival epithelial cells. Following treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL, the culture supernatant of human conjunctival epithelial cells promoted neutrophil migration and induced increases in the staining intensity of MPO and CitH3. In addition, increased NETs triggered the apoptosis of human conjunctival epithelial cells and IL-6, TNF-α, IFN-γ and IL-8 secretion in the culture supernatant of human conjunctival epithelial cells.
CONCLUSIONS
CDM induces human conjunctival epithelial cell damages, thereby promoting neutrophil migration and NETs formation, while the release of NETs further aggravates human conjunctival epithelial cell damages.
Animals
;
Humans
;
Extracellular Traps
;
Neutrophils
;
Interleukin-8/metabolism*
;
Dermatophagoides farinae
;
Interleukin-6/metabolism*
;
Tumor Necrosis Factor-alpha/metabolism*
;
Epithelial Cells
;
Interferon-gamma/metabolism*
;
Tetradecanoylphorbol Acetate/pharmacology*
5.Exploration and application of "internet + mobile medicine" in the process management of standardized residency training of obstetrics and gynecology
Meili PEI ; Li WANG ; Juan ZHAO ; Ting YANG ; Ruifang AN ; Xiaofeng YANG
Chinese Journal of Medical Education Research 2023;22(1):133-138
Objective:To assess the application effect of "internet + mobile medicine" on the process management of standardized residency training of obstetrics and gynecology.Methods:A total of 43 teachers including 14 administrators and 29 instructors who were not directly responsible for management, and 41 residents who participated in the standardized training in our base were enrolled as research objects. The data of process management were sorted by internet technology, and combined with personal wishes and casting lots, the residents were divided into two groups: the control group ( n=21) adopted traditional way while the observation group ( n=20) used mobile medicine for process management. By conducting multi-dimensional questionnaire and data comparison, we analyzed the evaluation of teachers and residents, the scores of different stage of residents, and the satisfaction of cervical cancer postoperative patients to medical care under different process management mode. SPSS 22.0 software was used for t test and chi-square test. Results:The application of "internet + mobile medicine" in standardized residency training obtained positive evaluation both by teachers and residents, and the ratio of considering "necessary" to reform the traditional residency training management mode in residents [95.1%(39/41)] and administrators [92.9%(13/14)] were both higher than that in instructors [62.1%(18/29)]. And 78.6%(11/14) of the administrators believed that cloud data management took less time. After 6 months of process management by different modes, the clinical diagnosis and treatment ability and skill operation ability scores of observation group were higher than those of control group [(93.6±5.6) vs. (89.4±5.7); (89.6±8.8) vs. (84.0±8.7)]. The "overall medical satisfaction rate" of cervical cancer patients was relatively high in both groups [100%(30/30) vs. 93.3%(28/30)], and the "very satisfied rate" of patients in the mobile medical group was higher than that in the traditional group [86.7%(26/30) vs. 63.3%(19/30)].Conclusion:The application of "internet + mobile medicine" is conducive to strengthening the process management and optimizing the quality of standardized residency training management of obstetrics and gynecology.
6.Incidence of maternal sepsis in ICUs of hospitals in Beijing: a multicenter cohort study
Zhiling ZHAO ; Jianan ZHANG ; Jianxin ZHANG ; Meili DUAN ; Jingjing XI ; Gaiqi YAO ; Yangyu ZHAO ; Qinggang GE ; Shining BO ; Qingtao ZHOU
Chinese Critical Care Medicine 2023;35(3):305-309
Objective:To investigate the epidemiological data of maternal sepsis in intensive care unit (ICU), analyze the common causes, outcomes of maternal sepsis, and the risk factors of multi-drug resistant (MDR) bacteria.Methods:A retrospective cohort study. Maternal sepsis cases admitted to ICUs of Peking University Third Hospital, Beijing Chao-Yang Hospital Affiliated to Capital Medical University, and Beijing Friendship Hospital Affiliated to Capital Medical University from January 2008 to September 2022 were enrolled. The following data were recorded: demographic characteristics, sequential organ failure assessment (SOFA) during infection, infection time, infection sites, invasive intervention measures before infection, microbial culture results, blood routine test during infection, body temperature, and clinical outcomes caused by infection. According to the time of sepsis occurrence, the patients were divided into pre-ICU sepsis group and ICU sepsis group, and the causes of sepsis in the two groups were analyzed. According to whether MDR occurred, the patients were divided into MDR group and non-MDR group, and clinical outcomes were analyzed. Multivariate Logistic regression was used to analyze the risk factors of MDR bacteria infection in obstetrics with sepsis.Results:160 patients were enrolled, among which 104 cases of sepsis happened before ICU and 56 cases of sepsis happened during ICU, 53 cases were with MDR bacteria and 107 cases were without MDR bacteria. The median age of the patients was 30.5 (28.0, 34.0) years old, the median temperature was 38.8 (38.2, 39.5) ℃, and the median white blood cell count (WBC) was 17.2 (13.2, 21.3)×10 9/L, the median SOFA score was 5.0 (3.0, 8.0), and 130 cases (81.2%) were referred from other hospitals. The main infection sites were uterine cavity in 64 cases (40.0%), lung in 48 cases (30.0%), abdominal and pelvic cavity in 30 cases (18.8%), urinary system in 27 cases (16.9%). Sepsis led to hysterectomy in 6 cases (3.8%), stillbirth in 8 cases (5.0%), and neonatal death in 2 cases (1.3%). The main surgical intervention measures were cesarean section (44 cases, accounting for 27.5%), followed by exploratory laparotomy (19 cases, 11.9%). The median length of ICU stay was 5.0 (3.0, 10.0) days, and the median hospital length was 14.0 (10.0, 20.8) days. Intrauterine infection was the primary cause of sepsis happened during ICU, accounting for 50.0% (28/56), of which postpartum hemorrhage accounted for 85.7% (24/28). The proportion of diabetes [28.3% (15/53) vs. 14.0% (15/107)], intrauterine operation [41.5% (22/53) vs. 23.4% (25/107)], intrauterine infection [50.9% (27/53) vs. 34.6% (37/107)] and bacteremia [18.9% (10/53) vs. 2.8% (3/107)] in the MDR group were significantly higher than those in the non-MDR group (all P < 0.05). Multivariate Logistic regression analysis showed that diabetes [odds ratio ( OR) = 2.348, 95% confidence interval (95% CI) was 1.006-5.480, P = 0.048] and intrauterine operation ( OR = 2.541, 95% CI was 1.137-5.678, P = 0.023) were independent risk factors for MDR bacterial infection in obstetrics with sepsis. Conclusions:Intrauterine infection is the common cause of maternal sepsis in ICU, and postpartum hemorrhage is the common cause of secondary intrauterine infection in ICU. MDR bacteria can lead to serious clinical outcomes. Diabetes and intrauterine operation are independent risk factors for MDR bacteria' infection.
7.Role and mechanism of miRNAs in alcoholic liver injury in rats
Meili CONG ; Tao LIU ; Xuan FEI ; Bei ZHOU ; Jianxin SUN ; Xiaoguo ZHAO
Chinese Journal of Comparative Medicine 2023;33(12):34-41
Objective To investigate the role and mechanism of miRNAs in alcoholic liver injury in rats.Methods Thirty male SD rats were randomly divided into model and control groups.The model group was gavaged with 56%liquor and the control group was gavaged with distilled water for 8 weeks.Liver tissue was collected,miRNAs were analyzed,and target genes of differentially expressed miRNAs were predicted by a rat miRNA chip.Gene ontology(GO)and KEGG pathway enrichment analysis were used to understand the function of differentially expressed miRNA target genes.A differentially expressed miRNA-mRNA-pathway regulatory network was constructed using Cytoscape to further screen important regulatory miRNAs versus important pathways.RT-qPCR was performed for selected miRNAs to validate the expression analysis.Results Twelve differentially expressed miRNAs(P<0.05,Fold change≥2)were screened out,including two upregulated and 10 downregulated miRNAs by comparative analysis of microarray data between model and control groups.GO classification annotation of differential miRNA target genes showed close associations between differentially expressed miRNAs and biological functions such as signal transduction,metabolic processes,antioxidant activity,cell killing,enzyme regulatory activity and biological regulation.Differentially expressed miRNA target genes in KEGG pathway analysis revealed that the AMPK signaling pathway,PI3K-Akt signaling pathway,Hippo signaling pathway,Wnt signaling pathway,cancer,autophagy,insulin resistance,Ras signaling pathway,and other signaling pathways might play major regulatory roles in alcoholic liver injury lesions.Hub miRNAs and pathways screened by constructing the differentially expressed miRNA-mRNA-pathway regulatory network were miR-145-5p,miR-107-3p,miR-297,Hippo signaling pathway,cancer,PI3K-Akt signaling pathway,and AMPK signaling pathway.qRT-PCR validated the gene expression trends,and gene chip result were consistent.Conclusions We established an miRNA profile of alcoholic liver injury in rats,which suggests that miR-145-5p,miR-107-3p,and miR-297 play major roles in the process of alcoholic liver pathology.
8.Glycyrrhizic acid⁃mediated protective effect of miRNAs on alcoholic liver inj ury in rats
Meili Cong ; Mayire Nuermaimaiti ; Bei Zhou ; Ruihua Chen ; Zhong Wang ; Huhu Zhu ; Xiaoguo Zhao
Acta Universitatis Medicinalis Anhui 2023;58(8):1374-1379
Objective :
To investigate the mechanism of miRNAs in glycyrrhizic acid treatment of alcohol⁃induced
liver injury in rats .
Methods :
45 male SD rats were randomly divided into glycyrrhizin group , model group and control group . The rats in glycyrrhizin group were given 56% liquor and glycyrrhizin , the rats in model group were
given 56% liquor , and the rats in control group were given distilled water for 8 weeks . The blood was collected and the serum was separated by centrifugation to detect the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) . RNA were extracted from liver tissues , miRNAs were detected by rat miRNA microarray , and their expression levels were analyzed . The miRNA target genes of differential miRNA were predicted . Gene Ontology (GO) and KEGG Pathway enrichment analysis were used to understand the function of differential miRNA , and the differential miRNA⁃mRNA⁃Pathway regulatory network was constructed using Cytoscape to further screen the regulatory key miRNA and key pathways . qRT⁃PCR was used to verify the expression of selected miRNA .
Results :
Compared with the model group , the glycyrrhizin group could significantly improve the liver tissue lesions , and reduce the liver serum AST and ALT levels (P < 0. 05) . Compared the microarray data of glycyrrhizin group and model group , a total of 13 differentially expressed miRNA were screened ( P < 0. 05 , fold change ≥ 1 . 5) , of which 10 were up⁃regulated and 3 were down⁃regulated . The GO classification annotation of differential miRNA target genes showed that differential miRNA were related to cell adhesion , antioxidant activity , metabolic process , biological process regulation , cell killing , immune system and other functions . The KEGG Pathway analyling pathway , Hippo signaling pathway , PI3K⁃Akt signaling pathway , wnt signaling pathway , apoptosis and other signaling pathways might play an important regulatory role in the improvement of alcoholic liver injury by glycyrrhic acid .
Conclusion
This study established the miRNA expression profile of glycyrrhizin in the treatment of alcoholic liver injury in rats , suggested that miR⁃615 , miR⁃107 ⁃3p and miR⁃292⁃5p might play an important role in the treatment of alcoholic liver injury by glycyrrhizin .
9.Exploration and study on the modern distance education in traditional Chinese medicine colleges and universities
Zhiqin GUO ; Yang YANG ; Hailing ZHAO ; Meili ZHAO
Chinese Journal of Medical Education Research 2022;21(10):1285-1287
This article takes the modern distance education of Beijing University of Chinese Medicine for example, and analyzes the current situation and future development paths of modern distance education in traditional Chinese medicine (TCM) colleges and universities, which has provided references for other TCM colleges and universities to set up modern distance education, and laid the foundation for further improving the quality of distance education and social influence, optimizing the modern TCM talent cultivation system, and enhancing the quality of TCM talent cultivation.
10.Intestinal microbiome changes in patients with postpartum depression
Zai YANG ; Yudan ZHANG ; Xin MU ; Meili PEI ; Yuan GAO ; Yajuan FAN ; Binbin ZHAO ; Xiancang MA
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(6):879-884
【Objective】 To analyze the changes of gut microbes in patients with postpartum depression so as to explore the relationship between postpartum depression and gut microbes. 【Methods】 A total of 60 postpartum subjects were recruited to participate in this study. The depression status of the participants was scored using the Edinburgh Postpartum Depression Scale (EPDS). Those with a score ≥13 were included in the postpartum depression group (PPD group), while those with a score less than 13 were included in the postpartum healthy control group (PPHC group). The feces of these 60 subjects were collected, and the fecal whole genome DNA was extracted for 16S rDNA sequencing. The data of changes in the bacterial diversity between the groups were obtained, and the possible correlation between the changes of intestinal microbes and postpartum depression was analyzed. 【Results】 The number of microorganisms in PPD patients was significantly reduced (P<0.001); the Chao1 index (P<0.001) and ACE index (P<0.001) of α diversity decreased significantly. There were also significant differences in β diversity between the two groups. Analysis of the bacteria in the groups showed that Acetanaerobacterium, Adlercreutzia, Allobaculum, Alloprevotella, Bifidobacterium, Christensenella, Escherichia, Eubacterium, Faecalicatena, Fusobacterium, Haemophilus, Intestinimonas, Lactobacillus, Megamonas, Monoglobumus, Muribaculum, Oscillospira, Paraprevotella, Streptococcus, Raoultibacter, Ruminococcus and Stomatobaculum were significantly enriched in PPHC group. In contrast, Kineothrix, Lachnoclostridium, Acinetobacter, Aquisphaera, Enterococcus, and Mucispirillum were enriched in PPD group. RDA/CCA analysis showed that EPDS was positively correlated with Prevotella, Kineothrix, and Alistipes, but negatively correlated with Lachnospira. 【Conclusion】 This study found that the intestinal flora of patients with postpartum depression was significantly disrupted, and there was a correlation between the intestinal flora and postpartum depression symptom score. Therefore, intestinal microbial markers may contribute to the diagnosis and treatment of patients with postpartum depression.


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