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.Mechanism of Buzhong Yiqitang Intervening in Pyroptosis of AIT in NOD.H-2h4 Mice Based on NLRP3/Caspase-1/GSDMD Pathway
Zhimin WANG ; Xiao YANG ; Yiran CHEN ; Ziyu LIU ; Qingyang LIU ; Chenghan GAO ; Chen WANG ; Tianshu GAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(12):8-15
ObjectiveTo explore the mechanism of Buzhong Yiqitang on pyroptosis in autoimmune thyroiditis (AIT) mice based on the NOD-like receptor hot protein domain related protein 3 (NLRP3)/cysteinyl aspartate specific proteinase-1(Caspase-1)/Gasdermin D (GSDMD) pathway. MethodSixty NOD.H-2h4 mice were divided into normal group, model group, low, medium, and high dose groups (4.10, 8.19, 16.38 g·kg-1)of Buzhong Yiqitang, and selenium yeast tablet group (0.26 mg·kg-1), with 10 mice in each group. Except for the normal group, all other groups were given 0.05% NaI by gavage for eight weeks to establish a model and then received the drug treatment for eight weeks. The serum levels of thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (TgAb) were detected using enzyme-linked immunosorbent assay (ELISA) method. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in mouse thyroid tissue. The immunohistochemical method was used to detect the protein expression of NLRP3, Caspase-1, interleukin-1β (IL-1β), and interleukin-18 (IL-18). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of NLRP3, Caspase-1, IL-1β, and IL-18. Western blot was used to detect the levels of pyroptosis-related proteins in thyroid tissue. ResultCompared with the normal group, the serum levels of TPO-Ab and TgAb in the model group were significantly increased (P<0.01). Thyroid follicles either increased in a cubic shape or were damaged and atrophied, with a large number of lymphocytes infiltrating around the follicles. Compared with the model group, the levels of TPO-Ab and TgAb in other groups were significantly reduced (P<0.01), and the morphology and structure of follicles were improved. The degree of lymphocyte infiltration was reduced. Among them, the medium dose group of Buzhong Yiqitang had the most significant reduction and improvement effect. Compared with the normal group, the positive products and mRNA expression of NLRP3, Caspase-1, IL-1β, and IL-18 proteins in the thyroid tissue of the model group significantly increased (P<0.01), and the protein expression levels of NLRP3, cleaved Caspase-1, IL-1β, IL-18, and GSDMD-N were significantly increased (P<0.05, P<0.01). Compared with the model group, the positive products and mRNA expression of NLRP3, Caspase-1, IL-1β, and IL-18 proteins in other groups were significantly reduced (P<0.05, P<0.01), with the most significant reduction effect in the medium dose group of Buzhong Yiqitang. The protein expression levels of NLRP3, cleaved Caspase-1, IL-1β, IL-18, and GSDMD-N were significantly reduced (P<0.05, P<0.01). ConclusionBuzhong Yiqitang can improve AIT, and its mechanism may be achieved by regulating the NLRP3/Caspase-1/GSDMD signaling pathway to inhibit pyroptosis.
3.Improvement of Thyroid Injury in AIT Mice by Inhibiting Ferroptosis Through Regulation of Nrf2/PPARγ/GPX4 Pathway by Buzhong Yiqitang
Ziyu LIU ; Zhuo ZHAO ; Yiran CHEN ; Huimin CAO ; Si CHEN ; Zhimin WANG ; Tianshu GAO ; Xiao YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):10-18
ObjectiveTo investigate the mechanism of Buzhong Yiqitang in ameliorating ferroptosis in autoimmune thyroiditis (AIT) mice based on the nuclear factor erythroid 2-related factor 2 (Nrf2)/peroxisome proliferator-activated receptor γ (PPARγ)/glutathione peroxidase 4 (GPX4) pathway. Method120 SPF-grade 7-8-week-old NOD.H-2h4 mice were randomly divided into control group, model group, low-, medium-, and high-dose Buzhong Yiqitang groups, and western medicine group, with 20 mice in each group. Except for the control group, all mice were fed with classic high-iodine water (0.05% NaI) to induce AIT models after 8 weeks. The low-, medium-, and high-dose Buzhong Yiqitang groups were administered 4.78, 9.56, 19.12 g·kg-1 of Buzhong Yiqitang, respectively, via gavage. The western medicine group was given 3.033×10-5 g·kg-1 selenium yeast tablet suspension via gavage, while the control and model groups were given an equal volume of distilled water via gavage. After 8 weeks of continuous treatment, samples were collected. The pathological morphology of mouse thyroid tissue was observed through hematoxylin-eosin (HE) staining,the content of serumantithyroid peroxidase autoantibody(TPOAb)and anti-thyroglobulin antibodies(TGAb)was measured by enzyme-linked immunosorbent assay (ELISA),the kit was used to detect the levels of superoxide dismutase (SOD), and malondialdehyde (MDA) in mouse serum. Immunofluorescence was used to detect the localized expression of GPX4 in thyroid tissue. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of Nrf2, PPARγ, solute carrier family 7 member 11 (SLC7A11), solute carrier family 3 member 2 (SLC3A2), lysolipid lecithin acyltransferase 3 (LPCAT3), and GPX4 mRNA in thyroid tissue. Western blot was used to detect the expression of Nrf2, PPARγ, SLC7A11, SLC3A2, LPCAT3, and GPX4 proteins in thyroid tissue. ResultCompared with control group, model group under light microscopy showed significant lymphocyte infiltration in the thyroid tissue, significantly increased levels of TGAb and TPOAb in serum (P<0.01), significantly increased MDA levels and decreased SOD levels in serum (P<0.01), significantly decreased expression of Nrf2, PPARγ, SLC7A11, SLC3A2, and GPX4 (P<0.01) in thyroid tissue, while the expression of LPCAT3 was significantly increased (P<0.01). Compared with model group, the Buzhong Yiqitang groups and the western medication group under light microscopy showed lymphocyte infiltration in the thyroid tissue of was decreased, significantly decreased levels of TPOAb and TGAb in serum (P<0.05,P<0.01), decreased MDA levels and increased SOD levels in serum(P<0.05,P<0.01),significantly increased expression of Nrf2, PPARγ, SLC7A11, SLC3A2, and GPX4, while the expression of LPCAT3 was significantly decreased (P<0.05,P<0.01) in the thyroid tissue. Compared with western medication group, Buzhong Yiqitang groups showed significant overall trends in the expression of Nrf2, PPARγ, SLC7A11, SLC3A2, GPX4, and LPCAT3 (P<0.05,P<0.01). ConclusionBuzhong Yiqitang can effectively improve the inflammatory injury of AIT, and its mechanism of action may be related to the regulation of Nrf2/PPARγ/GPX4 to inhibit ferroptosis.
4.Mechanism of Buzhong Yiqitang in Improving Autoimmune Thyroiditis by Regulating Th17 Cells Through miR-155/Ndfip1/Pten Axis
Xiaohui LI ; Zhuo ZHAO ; Yiran CHEN ; Huimin CAO ; Si CHEN ; Zhimin WANG ; Tianshu GAO ; Ziyu LIU ; Xiao YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):19-26
ObjectiveTo explore the mechanism of Buzhong Yiqitang in improving autoimmune thyroiditis (AIT) by regulating helper T cell 17(Th17) cells through microRNA-155 (miR-155)/Nedd4 family interaction protein 1 (Ndfip1)/phosphatase and tensin homology (Pten) axis. MethodThe 100 SPF grade 8 week-old NOD.H-2h4 mice were fed with high iodine water (0.05% NaI) for 8 weeks, and AIT model was made. They were divided into model group, Buzhong Yiqitang low-,medium-,and high-dose groups (4.78,9.56,19.12 g·kg-1·d-1) and selenium yeast tablet group (3.033×10-5 g·kg-1) according to random number table method. There were 20 mice in each group and 20 mice in the control group. The control group and the model group were given the same amount of distilled water. After 8 weeks of continuous administration, Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of miR-155-5p, Ndfip1, Pten, protein tyrosine kinase 1 (Jak1), signaling and transcriptional activator 3 (Stat3) retinoic acid-associated orphan receptor γt (RORγt), and interleukin-17 (IL-17) mRNA in mouse thyroid tissue. Western blot was used to detect the expression of Ndfip1, Pten, Jak1, Stat3, RORγt, and IL-17 proteins in mouse thyroid tissue, immunohistochemical method was used to detect the expression of Ndfip1 and Pten proteins in mouse thyroid tissue; flow cytometry was used to detect the proportion of Th17 cells in mouse spleen. ResultCompared with the control group, the proportion of Th17 cells was increased (P<0.01). The expressions of miR-155-5p, Jak1, Stat3, RORγt and IL-17 were increased (P<0.01), while the expressions of Ndfip1 and Pten were decreased (P<0.01). Compared with model group, the proportion of Th17 cells was decreased (P<0.05,P<0.01). The expressions of miR-155-5p, Jak1, Stat3, RORγt and IL-17 were decreased (P<0.05,P<0.01), while the expressions of Ndfip1 and Pten were increased (P<0.05,P<0.01). ConclusionThe application of Buzhong Yiqitang can improve the autoimmune disorder of AIT mice, the mechanism of which may be related to the regulation of Ndfip1/Pten axis by miR-155 and then the regulation of Th17 cell differentiation.
5.Effect of Buzhong Yiqitang on Fas/FADD/Caspase-8 Cell Apoptotic Signaling Pathway in Mice with Autoimmune Thyroiditis
Xiaohui LI ; Zhuo ZHAO ; Yiran CHEN ; Huimin CAO ; Si CHEN ; Zhimin WANG ; Tianshu GAO ; Ziyu LIU ; Xiao YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):27-34
ObjectiveTo investigate the effects of Buzhong Yiqitang on the tumor necrosis factor receptor superfamily member 6 (Fas)/Fas related death domain protein (FADD)/Caspase-8 cell apoptotic signaling pathway in mice model with autoimmune thyroiditis (AIT). MethodThere were 120 SPF grade NOD.H-2h4 mice aged 8 weeks, 100 of which were fed with high iodine water (0.05% NaI), and the AIT model was made after 8 weeks. According to random number table method, they were divided into model group, Buzhong Yiqitang low-dose, medium-dose and high-dose groups (4.78, 9.56, 19.12 g·kg-1), selenium yeast tablet group (3.033×10-5 g·kg-1), with 20 mice in each group and 20 control group. The apoptosis of thyroid cells was detected by in situ end labeling (TUNEL) after 8 weeks of administration. The real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of Fas, FADD, Caspase-8, and Caspase-3 in thyroid tissue. Western blot was used to detect the protein expression of Fas, FADD, Caspase-8, Caspase-3, cleaved Caspase-8, and cleaved Caspase-3 in thyroid tissue, and the immunohistochemical method was used to detect the protein expression of Fas and Caspase-3 in thyroid tissue. ResultCompared with control group, there were more positive expressions of apoptotic cells in model group under fluorescence microscope, and the expressions of Fas, FADD, Caspase-8, Caspase-3, cleaved Caspase-8 and cleaved Caspase-3 were significantly increased (P<0.05, P<0.01). Compared with model group, the positive expression of thyroid apoptotic cells in each administration group was decreased under fluorescence microscope, and the expressions of Fas, FADD, Caspase-8, Caspase-3, cleaved Caspase-8, cleaved Caspase-3 were significantly decreased (P<0.05, P<0.01). ConclusionBuzhong Yiqitang can effectively improve thyroid cell apoptosis and inflammatory injury in AIT mice. The mechanism may be related to the inhibition of Fas/FADD/Caspase-8 signaling pathway.
6.Analysis on the Preliminary Effect of Payment Reform in a Chinese Medicine Hospital Based on Interrupted Time Series
Xiaoyu ZHANG ; Tianshu CHU ; Lili WANG ; Limei JING
Chinese Health Economics 2024;43(9):29-34,39
Objective:In the background of the national promotion of DIP insurance payment reform,using empirical data to evaluate the initial effects of payment reforms in typical traditional Chinese medicine hospitals.Methods:It collected inpatient case data from a Chinese medicine hospital in S city,encompassing a total of 207 500 cases from 2016 to 2022,and the differences between before and after the reform were analyzed using key indicators such as medical costs.Interrupted time series was applied to analyze the initial effects of the reform based on three intervention time points:reform implementation,epidemic development and static management.Logistic regression was applied to explore the main influencing factors,and recommendations for refinement were made based on the results of the key informant interviews.Results:It was found that the percentage of the high hospitalization cost group,high self-pay cost group and high bed-day cost group increased after the reform.There was an overall increasing trend in bed-day costs after the reform.Age,being married,operation times and average grade of operations,and death were the common influences.Conclusion:The initial stage of the DIP insurance payment reform was not effective in regulating medical costs.Demographic changes,strong intervention in epidemics,and the tendency of Chinese medicine hospitals to perform surgeries all tend to lead to an increase in hospitalization costs.At the same time,factors such as the shortage of policy interpretation and the lack of incentives for medical staff have affected the effectiveness of the reform.It was suggested that the DIP insurance payment reform of the Chinese medicine hospitals should be further promoted by strengthening the interpretation of policies and changing the solidification model,establishing positive incentives for medical staff,and adding indicators of traditional Chinese medicine characteristics.This will further fully leverage the advantages of Chinese medicine characteristics and enhance the substance and quality of Chinese medicine services.
7.Constitution identification model in traditional Chinese medicine based on multiple features
Anying XU ; Tianshu WANG ; Tao YANG ; Xiao HAN ; Xiaoyu ZHANG ; Ziyan WANG ; Qi ZHANG ; Xiao LI ; Hongcai SHANG ; Kongfa HU
Digital Chinese Medicine 2024;7(2):108-119
Objective To construct a precise model for identifying traditional Chinese medicine(TCM)constitutions,thereby offering optimized guidance for clinical diagnosis and treatment plan-ning,and ultimately enhancing medical efficiency and treatment outcomes. Methods First,TCM full-body inspection data acquisition equipment was employed to col-lect full-body standing images of healthy people,from which the constitutions were labelled and defined in accordance with the Constitution in Chinese Medicine Questionnaire(CCMQ),and a dataset encompassing labelled constitutions was constructed.Second,heat-suppres-sion valve(HSV)color space and improved local binary patterns(LBP)algorithm were lever-aged for the extraction of features such as facial complexion and body shape.In addition,a dual-branch deep network was employed to collect deep features from the full-body standing images.Last,the random forest(RF)algorithm was utilized to learn the extracted multifea-tures,which were subsequently employed to establish a TCM constitution identification mod-el.Accuracy,precision,and F1 score were the three measures selected to assess the perfor-mance of the model. Results It was found that the accuracy,precision,and F1 score of the proposed model based on multifeatures for identifying TCM constitutions were 0.842,0.868,and 0.790,respectively.In comparison with the identification models that encompass a single feature,either a single facial complexion feature,a body shape feature,or deep features,the accuracy of the model that incorporating all the aforementioned features was elevated by 0.105,0.105,and 0.079,the precision increased by 0.164,0.164,and 0.211,and the F1 score rose by 0.071,0.071,and 0.084,respectively. Conclusion The research findings affirmed the viability of the proposed model,which incor-porated multifeatures,including the facial complexion feature,the body shape feature,and the deep feature.In addition,by employing the proposed model,the objectification and intel-ligence of identifying constitutions in TCM practices could be optimized.
8.Changes of lung function and inflammatory factors in rat models of coal workers' pneumoconiosis
Xiao HAN ; Tianshu WANG ; Jing SONG ; Linping WANG ; Huifang ZHANG ; Xiaoting LU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2024;42(8):561-567
Objective:To observe the changes of lung function and inflammatory factors in rat models of coal workers' pneumoconiosis at different time points.Methods:In June 2021, 96 healthy male SD rats with SPF grade were divided into 1, 3, and 6-month control group and dust staining group (coal dust group, coal silica dust group, quartz group) according to random number table method, with 8 rats in each group. After one week of adaptive feeding, a one-time non-exposed tracheal perfusion method (1 ml/ piece) was used. The dust dyeing group was given 50 g/L coal dust, coal silica mixed dust and quartz dust suspension, respectively, and the control group was given 0.9% normal saline solution. At 1, 3 and 6 months after perfusion, lung function was detected by animal lung function apparatus, then all lung tissues and alveolar lavage fluid were killed, and lung histopathological morphological changes were observed by HE staining, and the contents of interleukin (IL-1β), IL-18, IL-4 and IL-10 in alveolar lavage fluid were detected by ELISA. One-way analysis of variance was used to compare groups. Two factors (inter-group treatment factor (4 levels) and observation time factor (3 levels) ) were used in the analysis of the effects of inter-group treatment and treatment time on related indicators.Results:HE staining results showed that coal spot appeared in the lung tissue of coal dust group, coal spot and coal silicon nodule appeared in the lung tissue of coal dust group, and silicon nodule appeared in the lung tissue of quartz group. Compared with the control group, the forced vital capacity (FVC) and forced expiratory volume at 0.2 second (FEV 0.2) of rats in the dust staining group had interaction between the treatment and treatment time ( P<0.05). With the increase of dust dyeing time, FVC and FEV 0.2 decreased significantly at 3-6 months of dust dyeing, and the maximum gas volume per minute (MVV) decreased significantly at 1-3 months of dust dyeing ( P<0.05). The lowest lung function index was in quartz group, followed by coal-silica group and coal-dust group. There were statistically significant differences in the main effect and interaction effect of the pro-inflammatory factor IL-18 among all groups in treatment and treatment time (IL-18: F=70.79, 45.97, 5.90, P<0.001), and interaction existed. The highest content of inflammatory factors in alveolar lavage fluid of all dust groups was quartz group, followed by coal silica group and coal dust group. There were significant differences in the main effect and interaction effect of anti-inflammatory factors between groups and treatment time (IL-4: F=41.55, 33.01, 5.23, P<0.001, <0.001, <0.001; IL-10: F=7.46, 20.80, 2.91, P=0.002, <0.001, 0.024), and there was interaction. The highest content of anti-inflammatory factor was in quartz group, followed by coal silica group and coal dust group. Conclusion:Lung function decreased and levels of inflammatory fators increased in rat models of coal workers' pneumoconiosis, with the quartz group being the most severely damaged. Lung function is mainly impaired in thrid-six months, and the content of inflammatory factors begins to change in first-thrid months. MVV are the earliest and most obvious in lung function. IL-18 is suitable for monitoring changes in the pro-inflammatory response of coal workers' pneumoconiosis, and IL-10 is suitable for monitoring changes in anti-inflammatory response.
9.Sinicization and application on behavioral change technologies taxonomy v1(BCTTv1)
Xuefeng ZHONG ; Tianshu CHEN ; Xiaoting WANG
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(2):172-176
The first edition of the behavior change technologies taxonomy v1 (BCTTv1) is the most widely used classification of behavior change technologies, providing a series of shared and standardized terms to identify the effective components of behavioral interventions. This article translated BCTTv1 into Chinese, forming a Chinese version of BCTTv1. BCTTv1 consists of 93 behavior change techniques and they are divided into 16 groups according to a hierarchical structure. Each group contains 3-11 behavior change techniques, and each technique is named, defined, explained and illustrated with examples.Briefly describe the application of behavior change techniques in behavior intervention research, providing reference for promoting the application and promotion of behavior change intervention methods and techniques in China.
10.Value of serum KLF2 and NOS3 levels in the diagnosis and disease assessment of pa-tients with acute cerebral infarction of large-artery atherosclerosis type
Tianshu WANG ; Lijun JING ; Yaqiong YANG ; Junfang TENG
Chinese Journal of Arteriosclerosis 2024;32(6):527-531
Aim To investigate the levels of krüppel-like factor 2(KLF2)and endothelial nitric oxide synthase 3(NOS3)in the serum of patients with acute cerebral infarction(ACI)of large-artery atherosclerosis(LAA)type,and to analyze their value in the diagnosis and disease assessment of LAA type ACI.Methods A total of 150 patients with LAA type ACI were divided into mild group(n=36),moderate group(n=48),and severe group(n=66)based on their condition.Additionally,a control group(n=150)was selected from health exminers during the same period.The levels of serum KLF2 and NOS3 in each group were compared;receiver operator characteristic(ROC)curve was applied to analyze the diagnostic value of serum KLF2 and NOS3 levels for LAA type ACI and the predictive value for the occurrence of severe LAA type ACI,respectively.Results The serum KLF2 and NOS3 levels were significantly lower in LAA type ACI group than those in control group(P<0.05).The serum KLF2 and NOS3 levels in the mild,moderate and se-vere groups were significantly decreased in turn(P<0.05).The area under the curve(AUC)of the combined diagnosis of serum KLF2 and NOS3 for LAA type ACI was 0.858,with a sensitivity of 73.33%and a specificity of 86.00%,which was superior to the individual diagnosis of KLF2 and NOS3(Zcombined detection-KLF2=3.796,Zcombined detection-NOS3=4.689,all P<0.001).The AUC of combined prediction of serum KLF2 and NOS3 for the occurrence of severe LAA type ACI was 0.878,with a sensitivity of 77.27%and a specificity of 90.48%,which was superior to the independent prediction of KLF2 and NOS3(Zcombineddetection-KLF2=2.401,P=0.016;Zcombined detection-NOS3=3.070,P=0.002).Conclusions The serum levels of KLF2 and NOS3 in patients with LAA type ACI were significantly reduced and negatively correlated with the severity of the disease.The combination of the two has high evaluation efficacy in the diagnosis and disease prediction of LAA type ACI.

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