1.Preliminary study on the role of TM9SF2 knockdown in promoting the activity of the type I interferon signaling pathway to inhibit vesicular stomatitis virus replication.
Kang LI ; Xinyu WANG ; Ran YE ; Lingyun GUO ; Linxu WANG ; Nuo XU ; Tong ZHANG ; Xiaotao DUAN
Chinese Journal of Cellular and Molecular Immunology 2025;41(6):481-487
Objective To explore the effect of the knockdown of transmembrane 9 superfamily protein member 2 (TM9SF2) on the replication of vesicular stomatitis virus (VSV), and investigate its role in the mechanism of antiviral innate immunity. Methods Small interfering RNA (siRNA) was used to knock down the TM9SF2 gene in human non-small cell lung cancer A549 cells. The CCK-8 method was used to assess cell proliferation. A VSV-green fluorescent protein (VSV-GFP) infected cell model was established. The plaque assay was used to measure the viral titer in the supernatant. RT-qPCR and Western blotting were employed to quantify the mRNA and protein levels of VSV genome replication in A549 cells following VSV infection, as well as the expression of interferon β (IFN-β) mRNA and interferon regulatory factor 3 (IRF3) protein phosphorylation following polyinosinic-polycytidylic acid (poly(I:C)) stimulation. Results Compared to the negative control, the knockdown of TM9SF2 exhibited a significant effect, with no observed impact on A549 cell proliferation. The VSV-GFP infected A549 cell model was successfully established. After viral stimulation, fluorescence intensity was reduced following TM9SF2 knockdown, and the mRNA and protein levels of VSV were significantly downregulated. The viral titer of VSV was decreased. After poly(I:C) stimulation, TM9SF2 knockdown significantly upregulated the mRNA level of IFN-β and the phosphorylation level of IRF3 protein. Conclusion The knockdown of TM9SF2 inhibits the replication of vesicular stomatitis virus, and positively regulates the type I interferon signaling pathway, thus enhancing the host's antiviral innate immune response.
Humans
;
Virus Replication/genetics*
;
Signal Transduction
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Membrane Proteins/metabolism*
;
A549 Cells
;
Vesiculovirus/physiology*
;
Interferon-beta/metabolism*
;
Interferon Regulatory Factor-3/genetics*
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Interferon Type I/metabolism*
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Vesicular Stomatitis/immunology*
;
Gene Knockdown Techniques
;
Vesicular stomatitis Indiana virus/physiology*
;
RNA, Small Interfering/genetics*
2.Expression changes and clinical significance of IRAK1 and TRAF6 in patients with septic shock
International Journal of Laboratory Medicine 2024;45(3):341-346
Objective To investigate the expression changes and clinical significance of interleukin-1 recep-tor-associated kinase 1(IRAK1)and tumor necrosis factor receptor-associated factor 6(TRAF6)in patients with septic shock.Methods A total of 142 patients with septic shock admitted from November 2020 to No-vember 2022(septic shock group)were selected as the study subjects,and those who came to the hospital for physical examination during the same period were selected as the control group.Patients with septic shock were divided into survival group(100 cases)and death group(42 cases)according to their survival status after 28 days of hospitalization observation and treatment.The expression changes of IRAK1 and TRAF6 in pa-tients with septic shock were monitored at admission and after 2,4 and 6 days of treatment.The dynamic changes of acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score and sequential organ failure assessment(SOFA)score were recorded.Spearman correlation analysis was used to evaluate the correlation between IRAK1,TRAF6 and APACHE Ⅱ score and SOFA score in septic shock patients.The correlation be-tween IRAK1 and TRAF6 was analyzed by Pearson correlation analysis.Logistic regression was used to ana-lyze the factors influencing survival status of patients with septic shock.The diagnostic value of IRAK1 and TRAF6 in survival of patients with septic shock was analyzed by the receiver operating characteristic curve.Results The relative expression levels of IRAK1 and TRAF6 in septic shock group were significantly lower than those in control group,and APACHE Ⅱ score and SOFA score were significantly higher than those in control group,with statistical significance(P<0.05).Compared with admission,the relative expression levels of IRAK1 and TRAF6 in 2,4 and 6 days after treatment were significantly increased,while APACHE Ⅱ score and SOFA score were significantly decreased,with statistical significance(P<0.05).Compared with the death group,the relative expression levels of IRAK1 and TRAF6 in the survival group were higher at each corre-sponding time point,and the APACHE Ⅱ score and SOFA score were lower,with statistical significance(P<0.05).Correlation analysis showed that IRAK1 and TRAF6 were negatively correlated with APACHE Ⅱscores and SOFA scores in septic shock patients,while IRAK1 and TRAF6 were positively correlated(r=0.688,P<0.05).IRAK1,TRAF6 and APACHE Ⅱ scores were independent risk factors for survival of septic shock patients(P<0.05).The AUC of the combined diagnosis of IRAK1 and TRAF6 was significantly larger than that of IRAK1 alone(Z=2.044,P=0.041)and that of TRAF6 alone(Z=2.442,P=0.015).Conclusion The expression of IRAK1 and TRAF6 can evaluate the survival and prognosis of patients with septic shock.
3.Effect of LAG3 molecule on B lymphocyte subsets and its function in the liver of mice infected with Echinococcus multilocularis
Xu-Ran ZHENG ; Bing-Qing DENG ; Xue-Jiao KANG ; Yin-Shi LI ; Ainiwaer ABIDAN ; Qian YU ; Rousu ZIBIGU ; Duolikun ADILAI ; Mao-Lin WANG ; Hui WANG ; Chuan-Shan ZHANG ; Jing LI
Chinese Journal of Zoonoses 2024;40(6):529-536
This study was aimed at investigating the effect of lymphocyte activation gene-3(LAG3)on liver B lymphocyte subsets and their functions in WT and LAG3-KO mice infected with Echinococcus multilocularis(E.multilocularis).In a mouse model of E.multilocularis infection,the expression and localization of CD19 and α-SMA in liver were detected by immu nohistochemistry.CD80,CD86 and MHC-Ⅱ molecules expressed on B cells and their subsets in mice liver were detected by flow cytometry.After 12 weeks of infection,the area and percentage of CD19 in LAG3-KO group was slightly higher than that in WT group,but the difference was not statistically(t=-1.241、-1.237,P>0.05).The area and percentage of a-SMA in LAG3-KO group was higher than that in WT group(t=-3.224、-3.227,P<0.05).The proportion of CD80 and MHC-Ⅱ molecules expressed on liver B cells in LAG3-KO group was up-regulated(t=-2.379,-3.321,P<0.05).The percentage of liver B2 cells in LAG3-KO group was higher than that in WT group(t=-2.695,P<0.05).The expression of CD80 on Blb cells in LAG3-KO group was significantly up-regulated(t=-5.315,P<0.001).The proportion of CD80 of B2 cells in LAG3-KO group was lower than that in WT group(t=2.806,P<0.05).The expression of MHC-Ⅱ molecule in B2 cells in LAG3-KO group was up-regulated(t=-4.227,P<0.01).It is suggested that LAG3 molecules affected the B cell subsets and func-tion of mouse liver in the middle stage of E.multilocularis infection,especially B2 lymphocytes.LAG3 molecule exerted an in-hibitory effect on the activation of B cells and the expression of MHC-class Ⅱ molecules,suggesting that it may be involved in B cell exhaustion caused by E.multilocularis.
4.Application and effect of electronic signature verification technology in children's vaccination clinic
Jing YU ; Fengyang TANG ; Zhiguo WANG ; Lei ZHANG ; Li LIU ; Ran HU ; Jun GAO ; Bangmei DUAN ; Qing FAN ; Guodong KANG
Journal of Public Health and Preventive Medicine 2023;34(6):65-67
Objective To evaluate the application and effect of signature verification technology in children's vaccination clinics (CVC) of Jiangsu Province in 2020. Methods The signature verification data were derived from the Jiangsu Provincial Vaccination Integrated Service Management Information System, and the inquiry and registration, informed consent, vaccine traceability code scanning and observation information of children's vaccination clinics in different regions were analyzed. 210 doses of vaccination information were randomly selected from CVCs in each county, and the length of vaccination services in different regions was compared. Results During 2020, all of CVCs in Jiangsu were equipped with signature verification technology, and the signature verification rate of each vaccination sector was more than 99.90%. The length of outpatient vaccination service and overall length of stay in southern Jiangsu were slightly shorter than those in other regions. Conclusion The introduction of electronic signature verification technology in CVCs can effectively standardize the vaccination. It is necessary to expand the functions of electronic signature verification equipment, strengthen data analysis and utilization, and guide vaccination scientifically.
6.Distribution of monocyte subsets and their surface CD31 intensity are associated with disease course and severity of hemorrhagic fever with renal syndrome.
Xiaozhou JIA ; Chunmei ZHANG ; Fenglan WANG ; Yanping LI ; Ying MA ; Yusi ZHANG ; Kang TANG ; Ran ZHUANG ; Yun ZHANG ; Yan ZHANG
Chinese Journal of Cellular and Molecular Immunology 2023;39(5):439-444
Objective To investigate the relationship between disease courses and severity and monocyte subsets distribution and surface CD31 intensity in patients of hemorrhagic fever with renal syndrome (HFRS). Methods Peripheral blood samples from 29 HFRS patients and 13 normal controls were collected. The dynamic changes of classical monocyte subsets (CD14++CD16-), intermediated monocyte subsets (CD14++CD16+) and non-classical monocyte subsets (CD14+CD16++) and the mean fluorescent intensity (MFI) of CD31 on monocyte subsets were detected by multiple-immunofluorescent staining and flow cytometry. Results In acute phase of HFRS, the ratio of classical monocyte subsets to total monocytes was dramatically decreased compared to convalescent phase and normal control. It was still much lower in convalescent phase compared to normal controls. The ratio of classical monocyte subsets to total monocytes were decreased in HFRS patients compared to that in normal control, whereas there was no difference between severe/critical groups and mild/moderate groups. On the contrary, the ratio of intermediate monocyte subsets to total monocytes in acute phase of HFRS was significantly increased compared to convalescent phase and normal control. The ratio of intermediate monocyte subsets to total monocytes were increased in HFRS patients compared to that in normal control, whereas no difference was found between severe/critical groups and mild/moderate groups. Phases or severity groups had no difference in ratio of non-classical monocyte subsets to total monocytes. Additionally, the ratio of classical monocyte subsets had a tendency to decline and that of intermediate monocyte subsets showed an increase both to total monocytes between the acute and convalescent phases in 11 HFRS patients with paired-samples. Moreover, in acute phase of HFRS, the mean fluorescent intensity (MFI) of CD31 on three monocyte subsets all decreased, specifically classical monocyte subsets showed the highest MFI of CD31 while the normal control reported the highest MFI of CD31 in non-classical monocyte subsets. In convalescent phase, the MFI of CD31 on classical and intermediated monocyte subsets were both lower than that of normal control, while MFI of CD31 was still significantly lower than normal control on non-classical monocyte subsets. Finally, MFI of CD31 on classical and intermediated monocyte subsets in severe/critical group were both lower than those in mild/moderate group, showing no statistical difference in MFI of CD31 on non-classical monocyte subset across groups of different disease severity. Conclusion The ratio of classical and intermediated monocyte subsets to total monocytes are correlated with the course of HFRS, and so are the surface intensity of CD31 on these monocyte subsets with the disease course and severity. The surface intensity of CD31 on non-classical monocyte subsets, however, is correlated only with the course of the disease. Together, the underlying mechanisms for the observed changes in monocyte subsets in HFRS patients should be further investigated.
Humans
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Monocytes
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Lipopolysaccharide Receptors
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Hemorrhagic Fever with Renal Syndrome
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Receptors, IgG
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Disease Progression
7.A circRNA-miRNA-mRNA network analysis underlying pathogenesis of human heart failure.
Ran XU ; Jian WU ; Chun-Jie YANG ; Le KANG ; Yu-Yao JI ; Chang LI ; Zhi-Wen DING ; Yun-Zeng ZOU
Journal of Geriatric Cardiology 2023;20(5):350-360
BACKGROUND:
The molecular mechanisms of heart failure (HF) are still poorly understood. Circular RNA (circRNA) has been discovered in the heart in increasing numbers of studies. The goal of this research is to learn more about the potential roles of circRNAs in HF.
METHODS & RESULTS:
We used RNA sequencing data to identify the characteristics of circRNAs expressed in the heart and discovered that the majority of circRNAs screened were less than 2000 nt. Additionally, chromosomes One and Y had the most and least number of circRNAs, respectively. After excluding duplicate host genes and intergenic circRNAs, a total of 238 differentially expressed circRNAs (DECs) and 203 host genes were discovered. However, only four of the 203 host genes of DECs were examined in HF differentially expressed genes. Another study used Gene Oncology analysis of DECs host genes to elucidate the underlying pathogenesis of HF, and it found that binding and catalytic activity accounted for a large portion of DECs. Immune system, metabolism, and signal transduction pathways were significantly enriched. Furthermore, 1052 potentially regulated miRNAs from the top 40 DECs were collected to build a circRNA-miRNA network, and it was discovered that 470 miRNAs can be regulated by multiple circRNAs, while others are regulated by a single circRNA. In addition, a comparison of the top 10 mRNAs in HF and their targeted miRNAs revealed that DDX3Y and UTY were regulated by the most and least circRNA, respectively.
CONCLUSION
These findings demonstrated circRNAs have species and tissue specific expression patterns; while circRNA expression is independent on host genes, the same types of genes in DECs and DEGs worked in HF. Our findings would contribute to a better understanding of the critical roles of circRNAs and lay the groundwork for future studies of HF molecular functions.
8.Mediating effect of psychological resilience on self-efficacy and knee functional recovery in patients after artificial total knee arthroplasty
Chinese Journal of Practical Nursing 2023;39(28):2212-2217
Objective:To explore the mediating effect of psychological resilience on self-efficacy and knee functional recovery of patients after artificial total knee arthroplasty, and to provide theoretical basis for promoting functional recovery after artificial total knee arthroplasty.Methods:This was a cross-sectional survey study. A total of 250 patients undergoing artificial total knee arthroplasty at the Orthopedics Department of Beijing Jishuitan Hospital Affiliated to Capital Medical University were set as the research objects from September 2021 to March 2022, self-designed demographic questionnaire, General Self-Efficacy Scale (GSES), Connor Davidson Resilience Scale (CD-RISC) and Hospital for Special Surgery (HSS) scale were used to evaluate postoperative self-efficacy, psychological reselience and knee function.Results:A total of 225 valid questionnaires were recovered. The GSES score, CD-RISC score and knee function score of 225 patients were (26.45 ± 3.75), (68.01 ± 2.85) and (71.76 ± 4.53) points, respectively. Psychological resilience was positively correlated with self-efficacy and knee function score ( r=0.535, 0.675, both P<0.05), and self-efficacy was positively correlated with knee function score ( r=0.688, both P<0.05). Stratified regression analysis in showed that self-efficacy, tenacity, strength, optimism, and psychological resilience were included in the equation, adjusted for R2=0.812, indicating good fit of the model. The mediation model showed that psychological resilience was partially mediated between self-efficacy and functional recovery, with its mediation effects accounting for 25.49% of the total effect. Conclusions:The recovery of knee function in patients after artificial total knee arthroplasty is at a moderate level, and improving psychological resilience and self-efficacy may probably promote the recovery of knee function.
9.Multi-component comprehensive quality evaluation of Tongjing ointment
Ling-Chao RAN ; Jin-Ying FENG ; Zhi-Ming LI ; Kang-Kang JI
China Pharmacist 2023;26(11):319-331
Objective To establish a method for comprehensive quality evaluation of Tongjing ointment by multi-component quantification combined with chemometrics and grey correlation analysis(GRA).Methods With curcumin as the internal reference substance,HPLC-QAMS method was used to simultaneously determine the contents of limonin,evodiamine,rutaecarpine,bisdemethoxycurcumin,demethoxycurcumin,curcumin,6-gingerol,8-gingerol,10-gingerol,tetrahydropalmatine,dehydrocorydalin,dorydaline in Tongjing ointment.The quality of Tongjing ointment was evaluated by cluster analysis,principal component analysis,orthogonal partial least square discriminant analysis and GRA.Results The determination of 12 components manifested a good linear relationship in the range of mass concentration(r≥0.999 1).The average recovery was between 96.58%and 100.19%(RSD<2.0%,n=9).There were no significant difference between the measured value of external standard method and the calculated value of HPLC-QAMS(P>0.05).Tongjing ointment samples were classified into three eategories by chemometrics and it showed that curcumin,6-gingerol,bisdemethoxycurcumin,demethoxycurcumin,limonin and tetrahydropalmatine were the main potential markers affecting the quality of Tongjing ointment.GRA showed that the relative correlation degree was in the range of 0.317 3-0.624 8,and there were some differences in the quality of Tongjing ointment.Conclusion The established method can comprehensively evaluate the quality of Tongjing ointment.
10.Quality of life and influencing factors of postoperative accelerated rehabilitation of esophageal cancer patients.
Yuan YU ; Min LI ; Ran KANG ; Nuo Xiao Xuan WANG ; Xin Zhe LIU ; Qing Miao ZHU ; Jun CAO
Chinese Journal of Oncology 2022;44(4):341-346
Objective: To explore the quality of life and its influencing factors of enhanced recovery after surgery (ERAS) of esophageal cancer patients. Methods: The quality of life of 134 esophageal cancer patients was assessed using the quality of life assessment scale (EORTC QLQ-C30) developed by the European Cancer Research and Treatment Organization. Student's t test, One-way ANOVA and multiple linear regression statistical methods were used to analyze the effects of sociodemographic and clinical characteristics on patients' quality of life. Results: The overall score of quality of life (74.00) was lower than that of the general population (75.30). However, the scores of emotion and cognition in function dimension (93.97 and 95.77) were better than those of the general population (82.80 and 86.50). The results of fatigue, pain, insomnia and constipation in symptom dimension (14.18, 10.94, 11.69 and 5.72) were better than those of the general population (28.80, 20.50, 20.40 and 10.70). The pathological stage, body mass index and dietary were independent influencing factors for the quality of life of patients with esophageal cancer (P<0.05). Conclusions: ERAS can partially improve the quality of life of esophageal cancer patients. More attention should be paid to the esophageal cancer patients after surgery and take targeted measures to improve their quality of life.
Esophageal Neoplasms/surgery*
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Fatigue
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Humans
;
Quality of Life
;
Surveys and Questionnaires


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