1.Effect of Buzhong Yiqitang on Th17/Treg Immune Imbalance and Notch1 Signaling Pathway in AIT Mice
Zhuo ZHAO ; Nan SONG ; Ziyu LIU ; Pin LI ; Yue LUO ; Pengkun ZHANG ; Zhimin WANG ; Yuanping YIN ; Tianshu GAO ; Zhe JIN ; Xiao YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):19-27
ObjectiveTo explore the effect of Buzhong Yiqitang on the immune imbalance of helper T cell 17 (Th17)/regulatory T cell (Treg) and Notch1 signaling pathway in mice with autoimmune thyroiditis (AIT). MethodA total of 60 8-week-old NOD.H-2h4 mice were randomly divided into the normal group, model group, western medicine group (selenium yeast tablet, 32.5 mg·kg-1), and low-dose (4.78 g·kg-1·d-1), middle-dose (9.56 g·kg-1·d-1), and high-dose (19 g·kg-1·d-1) Buzhong Yiqitang groups, with 10 mice in each group. The normal group was fed with distilled water, and the other groups were fed with water containing 0.05% sodium iodide for eight weeks. After the animal model of AIT was formed spontaneously, the mice were killed under anesthesia after intragastric administration for eight weeks. Serum anti-thyroglobulin antibodies (TGAb), thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), and free thyroid hormone (FT4) were detected by enzyme-linked immunosorbent assay (ELISA), and thyroid tissue changes were observed by hematoxylin-eosin (HE) staining. The mRNA and protein expressions of retinoid-related orphan receptor-γt (RORγt), interleukin (IL)-17, forkhead box P3 (FoxP3), IL-10, Notch1, and hair division-related enhancer 1 (Hes1) in thyroid tissue were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultCompared with the normal group, the thyroid structure of the model group was severely damaged, and lymphocytes were infiltrated obviously. The levels of serum TGAb, FT3, and FT4 contents were significantly increased, and TSH content was significantly decreased (P<0.01). The mRNA and protein expression levels of RORγt, IL-17, Notch1, and Hes1 were significantly increased, while those of FoxP3 and IL10 were significantly decreased in the model group (P<0.01). Compared with the model group, thyroid structural damage and lymphocyte infiltration were improved in the treatment groups, and serum TGAb, FT3, and FT4 contents were significantly decreased. TSH content was increased, and mRNA and protein expression levels of RORγt, IL-17, Notch1, and Hes1 were decreased. mRNA and protein expression levels of FoxP3 and IL-10 were increased to different degrees (P<0.05, P<0.01), and the middle-dose Buzhong Yiqitang group had the most significant intervention effect. ConclusionBuzhong Yiqitang can alleviate the thyroid structural damage in AIT mice, and its mechanism may be related to improving the abnormal differentiation of Th17/Treg immune cells and inhibiting the activation of the Notch1 signaling pathway.
2.Analysis of cases of reinfection of past SARS-CoV-2 patients in Pudong New Area of Shanghai
Ge ZHANG ; Anran ZHANG ; Yilin JIA ; Li ZHANG ; Lipeng HAO ; Hongmei XU ; Yuanping WANG ; Chuchu YE ; Bo LIU ; Weiping ZHU ; Yixin ZHOU
Shanghai Journal of Preventive Medicine 2024;36(2):117-122
ObjectiveTo identify the rate, population characteristics, and vaccination history of repeat infections among previously infected people in the current epidemic based on the rate of repeat infection and population characteristics of different mutant strains at different times in Pudong New Area of Shanghai, and to provide reference for the prevention and control strategies of novel coronavirus repeat infections. MethodsA total of 9 250 investigated subjects were randomly selected from the new cases of asymptomatic infection and confirmed cases reported by Pudong New Area from March to May 2022. The investigation mainly focused on demographic characteristics, nucleic acid or antigen test results, and symptoms after infection. The repeat infection rates among different populations were compared, and logistic regression was used to analyze the impact of gender, age, and vaccination status on repeat infections. ResultsThe survey sample of 9 250 people had a response rate of 81.85%. There were 4 043 males (53.40%) and 3 528 females (46.60%), with a median age of 34 years old (P25, P75: 7, 61). The overall vaccine uptake rate was 59.44% (4 500/7 571). In December of 2022, there were 563 cases of repeat infection, with an infection rate of 7.44%. The lowest rate of repeat infection was seen in the 3‒ year-old group (2.86%) and the highest rate in the 30‒ year-old group (12.42%), with significant differences between different age groups. The repeated infection rate for those who had completed their vaccinations was significantly lower (6.57%) compared to those who had not (7.11%). The age groups of 3‒ years, 70‒79 years, as well as individuals who completed full vaccination and received booster shots were protective factors against repeat infections. ConclusionThe overall rate of reinfection among the infected in Shanghai during the spring of 2022 was low in the outbreak of the Omicron variant, and the rate of reinfection in the 3‒ year-old group was significantly lower than in other age groups. Completing the full course of vaccination significantly reduces the risk of reinfection. Although the reinfection rate is high in individuals who received booster shots, it remains a mitigating factor compared to those who do not receive the vaccine. It is recommended to continue monitoring reinfections in key populations and further strengthen immunization efforts.
3.Influence of NIC X-FILE and DENCO Pre-Shaper nickel-titanium instruments on the shaping of simulated curved canals
Yurong SHEN ; Rentong NAI ; Ling ZHAO ; Feigang LIU ; Caoyang YIN ; Yuanping GU ; Tieyi CHEN
Chinese Journal of Tissue Engineering Research 2024;28(3):387-391
BACKGROUND:Root canal preparation is a key step in root canal therapy.In recent years,with the rapid development of new nickel-titanium instruments,the results of root canal shaping ability have been significantly improved.However,the shaping abilities of different nickel-titanium instruments are also different.At present,there are few reports about the influence of the shaping ability of R-phase heat treatment NIC X-FILE and DENCO Pre-Shaper in simulated curved canals. OBJECTIVE:To compare the shaping effect of NIC X-FILE and DENCO Pre-Shaper nickel-titanium instruments in preparation for simulated curved canals. METHODS:Fifty simulated resin-curved canals were selected and randomly divided into two groups(n=25).NIC X-FILE and DENCO Pre-Shaper nickel-titanium files were used for root canal preparation.The root canal preparation time was recorded.Images of pre-and post-root canal preparation were taken.Image processing and analysis software were used to measure the amount of resin removed from the inner and outer walls of the root canal at each observation point.Statistical software was used to compare the root canal preparation time and centering ability of two kinds of mechanical nickel-titanium files. RESULTS AND CONCLUSION:(1)The average root canal preparation time was(1.58±0.02)minutes in the DENCO Pre-Shaper group and(2.22±0.03)minutes in the NIC X-FILE group.There was a significant difference between the two groups(P<0.05).(2)The amount of resin removed from the inner walls of the root canal of the DENCO Pre-Shaper group at 4,5,7,8,9,and 10 mm from the apical foramen was higher than that of the NIC X-FILE group(P<0.05).The amount of resin removed from the outer walls of the root canal of the DENCO Pre-Shaper group at 3,5,6,7,and 8 mm from the apical foramen was lower than that of the NIC X-FILE group(P<0.05).(3)At 2,4,5,6,7,8,and 9 mm away from apical foramen,the centering ability of the NIC X-FILE group was better than that of the DENCO Pre-Shaper group(P<0.05),and the centering ability of the NIC X-FILE group was the best at 4 mm away from apical foramen.(4)The results show that DENCO Pre-Shaper has higher mechanical efficiency than NIC X-FILE nickel-titanium root canal files.However,the centering ability of DENCO Pre-Shaper is inferior to NIC X-FILE nickel-titanium root canal files.
4.Mechanism of Buzhong Yiqitang in Ameliorating Thyroiditis Damage in AIT Mice by Modulating TLR4/NF-κB/AIM2 Signaling Pathway
Zhuo ZHAO ; Zhe JIN ; Zhengzheng LI ; Xuanlin GUO ; Jiayun LI ; Tongran GAO ; Pin LI ; Zhimin WANG ; Yuanping YIN ; Ziyu LIU ; Xiao YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):1-9
ObjectiveTo explore the mechanism of Buzhong Yiqitang in ameliorating inflammatory injury in autoimmune thyroiditis (AIT) mice based on the Toll-like receptor 4(TLR4)/nuclear transcription factor-kappa B(NF-κB)/absent in melanoma 2(AIM2)inflammasome signaling pathway. MethodThe 120 genetically susceptible 8-week-old NOD.H-2h4 mice were selected and randomly divided into control group, model group, low, medium and high dose groups of Buzhong Yiqitang (4.78, 9.56, 19.12 g·kg-1), and western medicine group (selenium yeast tablets, 3.033×10-5 g·kg-1). The AIT model mice in each group drank ad libitum 0.05% sodium iodide aqueous solution for 8 weeks to establish the AIT model, and the control group drank ad libitum distilled water. Eight weeks later, the mice in each dosing group were divided into groups and gavage. The swelling of thyroid tissue was observed with the naked eye, and the weight of spleen was weighed. The content of serum inflammatory factors interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) was measured by enzyme-linked immunosorbent assay (ELISA), and Real-time PCR was used to detect the expression of HMGB1, TLR4, AIM2, NF-κB p65,apoptosis-associated speck-like protein(ASC),cysteinyl aspartate-specific protease-1(Caspase-1), IL-1β mRNA. Western blot was used to detect the expression of high motility group protein 1 (HMGB1), TLR4, AIM2, NF-κB p65, phosphorylation(p)-NF-κB p65, ASC, Caspase-1, and IL-1β proteins in thyroid tissue, and immunofluorescence staining was used to observe the protein expression of HMGB1, AIM2, and NF-κB p65 in thyroid tissue of mice. ResultCompared with the control group, the thyroid tissue of mice in the model group was significantly swollen, the spleen quality was significantly increased, and the expression of HMGB1, TLR4, NF-κB p65, AIM2, ASC, Caspase-1, IL-1β in thyroid tissue was significantly increased (P<0.01). Compared with the model group, the swelling of thyroid tissue in mice in each dose group of Buzhong Yiqitang was improved, the quality of spleen was significantly reduced, and the expression of HMGB1, TLR4, AIM2, NF-κB p65, p-NF-κB p65, ASC, Caspase-1, IL-1β in thyroid tissue was significantly reduced (P<0.05, P<0.01). ConclusionBuzhong Yiqitang can effectively improve the inflammatory injury of AIT, and regulating the abnormal activation of the TLR4/NF-κB/AIM2 inflammasome signal pathway may be one of its intervention mechanisms.
5.Treating hyperhidrosis from liver and spleen
Mengzhen WANG ; Yue LUO ; Ziyu LIU ; Fengyi GUO ; Yuanping YIN ; Xiao YANG
Journal of Beijing University of Traditional Chinese Medicine 2024;47(4):490-495
Hyperhidrosis is a disease in which excessive sweat is secreted,resulting in an abnormal increase in systemic or local perspiration.In traditional Chinese medicine,hyperhidrosis belongs to the category of sweating disease.It is caused by an imbalance between yin and yang and abnormal excretion of body fluid.Many doctors treat the sweating disease from the perspective of heart,lung,and kidney;in contrast,we discuss the etiology and pathogenesis of hyperhidrosis from the perspective of liver and spleen,explain the significance of harmonizing the liver and spleen to improve the sweating disease,and put forward the key pathogenic factors of stagnation liver qi and spleen deficiency,and disharmony between nutrient qi and defensive qi,and the imbalance between yin and yang are the key pathogenic factor.Most of the clinical treatment start from the liver and spleen.The main treatment principle is based on soothing the liver and relieving depression,invigorating the spleen and nourishing blood,and regulating and harmonizing the nutrient qi and defensive qi,paying attention to the relationship between qi and blood in zangfu organs,invigorating the spleen and replenishing qi to consolidate its foundation,soothing the liver and relieving depression to regulate its qi,and invigorating qi and blood and perspiration.Treatment can be supplemented with Xiaoyao Powder plus-minus,mainly to strengthen liver wood and spleen soil,acquire nourishment,smooth qi,nourish the five zang,reconcile qi and blood,enhance physical strength and peace of mind,ensure sufficient qi to arrest sweating,coordinate the liver and spleen,balance the ascending and descending phases,and harmonize the five zang organs.
6.Study on the Mechanism of Panax Notoginsenosides in the Treatment of Sepsis Based on Network Pharmacology and Molecular Docking
Ping'e HUANG ; Ping YANG ; Wei HUANG ; Yuntao LIU ; Jin WANG ; Yuanping WANG ; Ye YE
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(7):1028-1034
Objective To explore the mechanism of Panax notoginsenosides in the treatment of sepsis based on network pharmacology and molecular docking technology.Methods The action targets of total saponins of Panax notoginsenosides were obtained by searching the databases of TCMSP,Swiss Target Prediciton and PharmMapper,and the disease-related targets of sepsis were searched in the databases of Genecard,Drugbank,Disgenet and OMIM.The selected targets of total saponins of Panax notoginsenosides were intersected with the disease-related targets of sepsis,which were used as potential targets for the treatment of sepsis.The protein-protein interaction(PPI)network of potential targets was constructed by STRING database,and the key targets were screened;the potential targets were analyzed by GO function and KEGG pathway enrichment analysis,and the drug-disease-target-pathway network was constructed;the molecular docking of five monomer saponins of Panax notoginsenosides and the key targets of Panax notoginsenosides in the treatment of sepsis was studied by AutoDock Vina software.Results A total of 206 potential targets of Panax notoginsenosides in the treatment of sepsis were obtained,and key targets such as AKT1,TP53,SRC,STAT3,JUN,TNF,IL6,MAPK1,PIK3R1 and IL1B were screened.A total of 2 548 biological process(BP)items,174 molecular function(MF)items and 47 cellular component(CC)items were obtained by GO functional enrichment analysis of potential targets,and 171 signal pathways were obtained by KEGG pathway enrichment analysis.The main active components of Panax notoginsenosides R1,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and ginsenoside Rd have strong binding activity with key targets AKT1,TP53,STAT3,SRC and JUN.Conclusion Panax notoginsenosides may act on the main signal pathways such as PI3K-Akt and AGE-RAGE through the key targets such as AKT1,TP53,SRC,STAT3 and TNF,and then affect the physiological processes such as inflammation,apoptosis and blood coagulation in sepsis,and play a role in the treatment of sepsis.
7.Potential Mechanism of Buzhong Yiqitang for Autoimmune Thyroiditis Based on miRNA Sequencing
Ziyu LIU ; Zhimin WANG ; Nan SONG ; Huimin CAO ; Si CHEN ; Yuanping YIN ; Tianshu GAO ; Xiao YANG
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(21):192-200
ObjectiveTo determine the influence of Buzhong Yiqitang on miRNA expression in thyroid tissues of mice with autoimmune thyroiditis (AIT). MethodThirty female 8-week-old NOD.H-2h4 mice were randomly assigned into normal control group, model group, and Buzhong Yiqitang group (BG), 10 in each group. Mice were subjected to a diet containing 0.05% sodium iodide for 8 weeks to build the AIT mouse model. After 8 weeks of administration (ig), samples were collected. A thyroid biopsy was performed on each group of mice, and differential miRNAs in thyroid tissues from each group of mice were analyzed based on experimental validation and bioinformatics. ResultCompared with the conditions of normal control group, thyroid lymphocytes had significant inflammatory infiltration, and there was an increase in serum TgAb level and interleukin(IL)-6 and IL-17 expression and a decrease in IL-1β expression in mice of the model group (P<0.05, P<0.01). In addition, 154 differentially expressed miRNAs were found. Compared with the conditions of model group, the degree of thyroid tissue inflammation was alleviated, and serum TgAb level, and IL-1β, IL-6 and IL-17 expression of mice treated with the Buzhong Yiqitang were reduced (P<0.05, P<0.01). Additionally, 112 differentially expressed miRNAs were identified in the BG group. Validation using real-time polymerase chain reaction(Real-time PCR) showed the same trend for miR-326-3p, miR-128-3p, miR-223-5p, miR-141-3p, miR-871-3p, and miR-204-3p as that obtained from miRNA sequencing. In particular, gene ontology(GO) functions were enriched for regulation of T cell activation, oxidative stress, and miRNA binding. Pathways identified by Kyoto encyclopedia of genes and genomes(KEGG)database tended to be enriched in phosphatidylinositol 3-kinases(PI3K)/protein kinase B(Akt), mitogen-activated protein kinase(MAPK), and cyclic adenosine monophosphate(cAMP) signaling pathways. Based on miRNA prediction differences, three key genes were identified: SMAD3, JAK2, and STAT3. ConclusionBushong Yiqitang might treat autoimmune thyroiditis by regulating 6 miRNAs.
8.The validity of the rehabilitation set of the International Classification of Functioning, Disability and Health in assessing aging-related disability
Shouguo LIU ; Juan JIN ; Xia ZHANG ; Juan YAN ; Mengqiu YE ; Yuanping ZHAO ; Hong XIE ; Jianan LI
Chinese Journal of Physical Medicine and Rehabilitation 2021;43(1):12-16
Objective:To explore the validity of a function assessing tool based on the International Classification of Functioning, Disability and Health′s (ICF′s) rehabilitation set in assessing aging-related disability.Methods:A total of 1610 elderly people from 15 nursing homes across China were assessed using the tool based on the ICF′s rehabilitation set and with the 12-item short form health survey (SF-12). The structural validity of the responses was analyzed using factor analysis, and criterion-related validity was also evaluated.Results:The factor analysis yielded three factors with eigenvalues greater than 1. Their cumulative explanatory power was 74.4%. Item d550 eating had double loading in the factor analysis. The item scores and the total scores of the disability assessment tool were significantly negatively correlated with the physiological function domain scores and the psychological function domain scores.Conclusion:The function assessment tool based on the ICF′s rehabilitation set when combined with a numerical rating scale has good structural and criterion-related validity in the assessment of disability due to aging.
9.Effects of m 6A methylase METTL14 mediated long-chain non-coding RNA EIF3J-AS1 on the migration and invasion of cholangiocarcinoma cells
Liu OUYANG ; Hao ZHENG ; Wenying CHENG ; Yuanping TAO ; Huifen LI ; Xiaoling LI ; Lei ZHANG ; Bo LI ; Shiwei GUO ; Xiangui HU ; Gang JIN
Chinese Journal of Pancreatology 2021;21(2):123-128
Objective:To investigate the effects of methyltransferase-like protein 14 (METTL14)-mediated long-chain non-coding RNA EIF3J antisense RNA1 (Inc EIF3J-AS1) on the migration and invasion of cholangiocarcinoma cells and its mechanism.Methods:From September 2017 to December 2018, 10 pairs of cholangiocarcinoma and adjacent normal tissues were collected from the First Affiliated Hospital of Naval Medical University, which were surgically resected and pathologically confirmed. The expression of METTL14 mRNA and Inc EIF3J-AS1 in cholangiocarcinoma tissues was detected by fluorescence quantitative PCR, and the protein expression of METTL14 was detected by Western blotting. Cholangiocarcinoma cell lines HUCCTI and RBE were divided into control group and METTL14 or Inc EIF3J-AS1 knockdown group. The corresponding normal lentivirus was transfected in the control group, and METTL14 or Inc EIF3J-AS1 knockdown group was transfected with lentivirus that interfered with the expression of METTL14 or Inc EIF3J-AS1, respectively. Transwell assay was used to detect the ability of cell migration and invasion, and Western blotting was used to detect the expression of epidermal growth factor receptor (EGFR) and AKT protein.Results:The expressions of METTL14 mRNA and lnc EIF3J-AS1 in cholangiocarcinoma tissues were significantly higher than those in adjacent normal tissues (0.075±0.012 vs 0.031±0.006, 0.140±0.032 vs 0.064±0.012), and there was a positive correlation between expression of METTL4 mRNA and expression of lnc EIF3J-AS1 ( r=0.883, P=0.0007). The expression of METTL14 protein in cholangiocarcinoma tissues was higher than that in adjacent normal tissues (0.354±0.131 vs 0.187±0.183). Compared with the control group, the expression of lnc EIF3J-AS1 was significantly lower in METTL14 or Inc EIF3J-AS1 knockdown group (0.217±0.020 vs 1.000±0.052, 0.149±0.066 vs 1.000±0.045). The migration and invasion ability of cell lines HUCCTI and RBE decreased significantly in lnc EIF3J-AS1 knockout group (5.00±0.58 vs 23.33±0.33, 20.33±0.67 vs 70.67±0.33; 12.00±0.58 vs 25.00±2.52, 22.33±0.89 vs 43.67±0.33). The expression of EGFR and p-AKT/AKT protein were also significantly decreased (0.109±0.015 vs 1.000±0.018, 0.226±0.036 vs 1.000±0.051; 0.118±0.052 vs 1.000±0.069, 0.132±0.098 vs 1.000±0.023). The above differences were statistically significant (all P<0.05). Conclusions:Abnormal expression of lnc EIF3J-AS1 in cholangiocarcinoma mediated by METTL14 can promote tumor cell migration and invasion.
10.Effects of miR-29b-3p targeting PTEN on congenital heart disease
Ping LIU ; Maiqiao YANG ; Yetian YANG ; Yiting WANG ; Lili LIU ; Yuanping MA ; Fang WANG ; Chuan HE
Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(6):868-874
【Objective】 To investigate the effects of miR-29b-3p on congenital heart disease and its mechanism. 【Methods】 The expression level of miR-29b-3p in serum from CHD patients and normal individuals, and in cells was detected by RT-qPCR. Dimethylsulfoxide (DMSO) was used to induce P19 cells to differentiate into cardiomyocytes. Western blot was used to measure the expression levels of cardiogenesis-associated genes, and phosphatase and tensin homolog (PTEN) level in cells. The proliferation and migration of cardiomyocytes were measured by CCK-8 and Transwell assay, respectively. Dual-luciferase gene reporter assay was used to verify the targeted relationship between miR-29b-3p and PTEN. 【Results】 Compared with that of normal individuals, the expression of miR-29b-3p in CHD patients was decreased. During differentiation, miR-29b-3p level was higher at late stage than that at early stage. Downregulated miR-29b-3p inhibited the differentiation of P19 cells into cardiomyocytes, and inhibited cell proliferation and migration. miR-29b-3p targeted PTEN. The increased PTEN level induced by miR-29b-3p knockdown inhibited the differentiation of P19 cells, and proliferation and migration of cardiomyocytes. 【Conclusion】 miR-29b-3p was downregulated in the serum of CHD patients. The downregulation of miR-29b-3p inhibited the differentiation of P19 cells, proliferation and migration of cardiomyocytes by targeting and regulating PTEN.

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