1.Impacts of galangin on immune inflammatory response in preeclampsia rats by regulating IL-6/STAT3 signaling pathway
Dongfang LIU ; Yi WANG ; Senji HAN ; Chong SUN ; Ningning SUN ; Tianshu LONG
Chinese Journal of Immunology 2024;40(12):2513-2517,2524
Objective:To investigate the impacts of galangin(Gal)on the immune inflammatory response in preeclampsia(PE)rats by regulating the IL-6/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods:Pregnant rats were injected subcutaneously with L-arginine methyl ester(L-NAME 125 mg/kg)to establish a PE model.After successful modeling,they were randomly grouped into Model group,low,medium and high doses Gal groups(10,30,and 60 mg/kg)and activator group(60 mg/kg Gal+0.05 mg/kg IL-6/STAT3 pathway activator rIL-6).Twelve pregnant rats of the same period were randomly selected as the control group,and each group was given corresponding drugs by gavage and intraperitoneal injection,once a day for a continuous week.The levels of mean caudal artery pressure(MAP)and 24-hour urinary protein and blood urea nitrogen(BUN)in rats during the third trimester of pregnancy were measured using a blood pressure tester and a fully automated biochemical analyzer;the number and weight of the average offspring of pregnant mice in each group were calculated,and the pregnancy outcome was analyzed;HE staining was applied to observe the pathological damage of rat placenta;ELISA was applied to detect the levels of IL-6,IL-1β and TNF-α in the placenta of rats in each group;the proportion of T lymphocytes in rat peripheral blood was detected by flow cytometry;Western blot was applied to detect the expression of IL-6/STAT3 pathway protein in rat placenta.Results:Compared with control group,the lev-els of MAP,24-hour urinary protein,BUN,IL-6,IL-1β,TNF-α,the CD8+T cells,and the expression levels of IL-6 and p-STAT3/STAT3 proteins in Model group were obviously increased,the placenta tissue was severely damaged,the mean offspring number and mean offspring weight,the proportions of CD3+T,CD4+T,and CD4+/CD8+T cells were decreased(P<0.05);compared with Model group,the levels of MAP,24-hour urinary protein,BUN,IL-6,IL-1β,TNF-α,the CD8+T cells,and the expression levels of IL-6 and p-STAT3/STAT3 proteins in Gal low,medium and high doses groups were obviously decreased,the damage to placental tissue was reduced,the mean offspring number and mean offspring weight,the proportions of CD3+T,CD4+T,and CD4+/CD8+T cells were in-creased(P<0.05);compared with Gal high-dose group,the levels of MAP,24-hour urinary protein,BUN,IL-6,IL-1β,TNF-α,the CD8+T cells,and the expression levels of IL-6 and p-STAT3/STAT3 proteins in activator group were obviously increased,the placenta tissue was aggravated,the mean offspring number and mean offspring weight,the proportions of CD3+T,CD4+T and CD4+/CD8+T cells were decreased(P<0.05).Conclusion:Gal may play a role in alleviating PE symptoms and improving pregnancy outcomes by inhibit-ing the IL-6/STAT3 pathway,inhibiting inflammatory reactions,and regulating cellular immunity.
2.Progress in clinical research of Claudin-18.2 targeted therapy for advanced gastric cancer
Yiyi YU ; Xiao WANG ; Tianshu LIU
Tumor 2024;44(1):34-43
Gastric cancer is one of the most common malignant tumors of the digestive system.Most gastric cancer patients are already in advanced stage when initially diagnosed.Advanced gastric cancer has a poor prognosis and has received limited benefit from chemotherapy-based therapies.In the recent years,with the development of biomolecular technologies,more and more researches have set their focus on the search for key molecules that affect the progression of gastric cancer,prognostic indicators,and therapeutic targets.Claudin-18.2 is a promising novel tumor therapeutic target which brings new hope for targeted therapy for gastric cancer.Various types of Claudin-18.2-targeting drugs have been developed,including monoclonal antibody,bispecific antibody(BsAb),antibody-drug conjugate(ADC),chimeric antigen receptor T cell(CAR-T),and so on.At present,several clinical trials have been carried out and the results are encouraging.This review summarizes the progress in clinical trials of Claudin-18.2 targeted therapy,in the hope to provide reference for improving the prognosis of patients with advanced gastric cancer.
3.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.
4.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.
5.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.
6.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.
7.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.
8.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.
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.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.

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