1.Comparison of the predictive performance of SARIMA, Prophet, and BSTS models in forecasting the incidence of hand, foot, and mouth disease
LU Wenhai ; KONG Xiaojie ; SONG Lixia ; LU Chunru ; YU Bikun ; XIE Yan
Journal of Preventive Medicine 2026;38(1):79-84
Objective:
To compare the predictive performance of the seasonal autoregressive integrated moving average (SARIMA) model, the Prophet model, and the Bayesian structural time series (BSTS) model in forecasting the incidence of hand, foot, and mouth disease (HFMD) , so as to provide a basis for optimizing the early warning system of this disease.
Methods:
Weekly incidence data of HFMD in Longgang District, Shenzhen City from 2014 to 2024 were collected. The HFMD incidence data from 2014-2019 and 2023 were used as the training set to construct SARIMA, Prophet, and BSTS models, while the data from 2024 were used as the test set to compare and evaluate the predictive performance of the three models. The technique for order preference by similarity to ideal solution (TOPSIS) method was employed to calculate the C-value. This approach integrates multiple evaluation metrics, such as the mean absolute error (MAE), mean squared error (MSE), root mean squared error (RMSE), and symmetric mean absolute percentage error (SMAPE), to comprehensively assess model performance.
Results:
A total of 150 111 cases of HFMD were reported in Longgang District from 2014 to 2024, with an average annual incidence of 400.72/105. The weekly incidence fluctuated between 0 and 63.78/105, exhibiting a bimodal seasonal pattern characterized by a primary peak from May to July and a secondary peak from September to October. In the training set, all three models demonstrated a good fit to the bimodal epidemic trend of HFMD, with the BSTS model achieving the best fit. The BSTS model yielded performance metrics as follows: MAE=0.124, MSE=0.050, RMSE=0.223, SMAPE=0.021, and a C-value of 1.000. In the test set, all three models, including SARIMA, Prophet, and BSTS, performed well for short-term predictions (≤16 weeks), with the Prophet model showing relatively superior predictive performance. However, the prediction accuracy of all models declined as the forecast horizon extended. During the primary peak period (May-July), the Prophet model exhibited better predictive performance, whereas the BSTS model performed relatively better during the secondary peak period (September-October).
Conclusions
For the short-term forecasting of weekly HFMD incidence, the Prophet model outperformed both the SARIMA and BSTS models. During the primary peak period, the Prophet model demonstrated superior predictive performance, whereas the BSTS model exhibited better accuracy in forecasting the secondary peak period.
2.Comprehensive Clinical Evaluation of Xiaojie'an Capsules in Treatment of Uterine Fibroids with Qi Stagnation and Blood Stasis Syndrome
Yutong MA ; Zhenkai LU ; Yanming XIE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):235-243
ObjectiveTo systematically organize the relevant research on the treatment of uterine fibroids with Xiaojie'an capsules through clinical evidence evaluation and comprehensive value analysis in the "6+1" dimension,highlighting the efficacy and characteristics of Xiaojie'an capsules. MethodsBased on evidence-based medicine,epidemiology,pharmacoeconomics,health technology assessment (HTA), and other methodologies,this study adopted qualitative and quantitative evaluation methods. Through a questionnaire survey,official website data information,published literature, and secondary evaluation of real world data,a total of "6+1" dimensions were used to build an evaluation system for the safety,effectiveness,economy,innovation,suitability,accessibility of Chinese patent medicines, and characteristics of traditional Chinese medicines. Results① Safety:From January 2009 to March 2023,the National Adverse Drug Reaction Monitoring Center reported 159 cases (248 cases) of general adverse reactions and four cases of serious adverse reactions to Xiaojie'an capsules,all of which were classified as general adverse reactions after expert judgment. Through literature search,a total of 6 865 patients were treated with Xiaojie'an capsules,of which 26 articles reported adverse reactions to Xiaojie'an capsules,resulting in a total of 244 adverse reactions. The clinical manifestations of adverse reactions to Xiaojie'an capsules include nausea,vomiting,stomach discomfort,diarrhea,rash,itching,dizziness,headache,insomnia,etc. The prognosis was good, and the drug surveillance system was sound. Based on the evaluation of known risks and sufficient evidence,in terms of known risk assessment, according to the monitoring results of the spontaneous reporting system (SRS), there were four cases of serious adverse reactions to Xiaojie'an capsules. After expert discussion, the four cases of serious adverse reactions were finally judged as general adverse reactions, and the known risks were considered to be small in combination with the known risk evaluation criteria. According to the types of studies carried out on Xiaojie'an capsules (randomized controlled trial and its systematic review, SRS data analysis, real world human experience studies, and non-clinical safety studies) and the evaluation criteria of sufficient evidence, it was considered that Xiaojie'an capsules had sufficient evidence. The safety evidence of this variety was sufficient, and the result was confirmed. ② Effectiveness: Meta-analysis results showed that the combination of Xiaojie'an capsules and Mifepristone tablets had better clinical efficacy compared to Mifepristone tablets alone and had a better effect in reducing the maximum uterine fibroid volume. Due to limitations and imprecision caused by downgrading factors,the quality of evidence was evaluated as Grade C. Based on comprehensive effectiveness evidence and value evaluation,the effectiveness evidence of this variety was relatively sufficient, and the results were clear ③ Economy:The economic evaluation results showed that treating one more patient with uterine fibroid required an additional cost of 5 438.57 yuan by using the combination of Xiaojie'an capsules and Mifepristone tablets,which was lower than the patient's willingness to pay (36 883 yuan according to 2022 National Bureau of Statistics data). This suggested that compared with using Mifepristone tablets alone,the combination of Xiaojie'an capsules and Mifepristone tablets had a certain economy. The sensitivity analysis results were relatively robust,indicating that this variety had good economy ④ Innovation:Currently,most traditional Chinese medicine treatments for uterine fibroids advocate the method of attacking evil,which can easily harm the body. However,Xiaojie'an capsules not only soften and disperse nodules but also balance supporting the body and supplementing deficiency. The comprehensive evaluation showed sufficient evidence of the innovation of this variety,and the results were confirmed. ⑤ Suitability: The service information of Chinese patent medicines derived from the drug was complete,and the questionnaire survey results showed that the drug was suitable for clinicians,nurses,pharmacists, and patients in multiple dimensions such as individual compliance,system, and management. The comprehensive evaluation of the suitability of the product had sufficient evidence and clear results. ⑥ Accessibility:The daily cost of Xiaojie'an capsules was 16.97 yuan,and the treatment cost was 407.16 yuan. It was affordable for urban residents and rural residents and was available for sale in 23 provinces, cities,municipalities,and autonomous regions across the country. The medicinal resources were abundant and sustainably supplied,and there was sufficient evidence of accessibility with clear results. ⑦ Characteristics of traditional Chinese medicine:Xiaojie'an capsules were derived from the classic Dai medicine formula of Dr. CHEN Benshan,a renowned Dai doctor,and they have been clinically used for over 20 years. Dai medicine originated from Xishuangbanna,Yunnan province,with the core theory of "four pagodas and five elements". The comprehensive evaluation showed that this variety exhibited significant characteristics of traditional Chinese medicine. ⑧ Comprehensive evaluation of clinical value:Based on the "6+1" dimensions mentioned above,combined with the multi-criteria decision analysis (MCDA) model and calculations using CSC v2.0 software,the results showed that there was sufficient evidence for the clinical value of Xiaojie'an capsules,and the results were clear. ConclusionThe comprehensive evaluation of Xiaojie'an capsules has sufficient evidence of safety and innovation,and the results are confirmed. The evidence of effectiveness,suitability,and accessibility is relatively sufficient,with clear results. The economy is good,and the characteristics of traditional Chinese medicine are prominent. The comprehensive evaluation of clinical value believes that there is sufficient evidence of the clinical value of Xiaojie'an capsules,and the results are clear. In the future,it is recommended to focus on clinical value, conduct high-quality and effective research that is in line with the characteristics of traditional Chinese medicine, and improve acute toxicity testing to provide a scientific and objective basis for clinical efficacy and safety research.
3.Molecular epidemiological characterization of influenza A(H3N2) virus in Fengxian District, Shanghai, in the surveillance year of 2023
Hongwei ZHAO ; Lixin TAO ; Xiaohong XIE ; Yi HU ; Xue ZHAO ; Meihua LIU ; Qingyuan ZHANG ; Lijie LU ; Chen’an LIU ; Mei WU
Shanghai Journal of Preventive Medicine 2025;37(1):18-22
ObjectiveTo understand the epidemiological distribution and gene evolutionary variation of influenza A (H3N2) viruses in Fengxian District, Shanghai, in the surveillance year of 2023, and to provide a reference basis for influenza prevention and control. MethodsThe prevalence of influenza virus in Fengxian District in the 2023 influenza surveillance year (April 2023‒March 2024) was analyzed. The hemagglutinin (HA) gene, neuraminidase (NA) gene, and amino acid sequences of 75 strains of H3N2 influenza viruses were compared with the vaccine reference strain for similarity matching and phylogenetic evolutionary analysis, in addition to an analysis of gene characterization and variation. ResultsIn Fengxian District, there was a mixed epidemic of H3N2 and H1N1 in the spring of 2023, with H3N2 being the predominant subtype in the second half of the year, and Victoria B becoming the predominant subtype in the spring of 2024. A total of 75 influenza strains of H3N2 with HA and NA genes were distributed in the 3C.2a1b.2a.2a.2a.3a.1 and B.4 branches, with overall similarity to the reference strain of the 2024 vaccine higher than that of the reference strain of the 2022 and 2023 vaccine. Compared with the 2023 vaccine reference strain, three antigenic sites and one receptor binding site were changed in HA, with three glycosylation sites reduced and two glycosylation sites added; where as in NA seven antigenic sites and the 222nd resistance site changed with two glycosylation sites reduced. ConclusionThe risk of antigenic variation and drug resistance of H3N2 in this region is high, and it is necessary to strengthen the publicity and education on the 2024 influenza vaccine and long-term monitoring of influenza virus prevalence and variation levels.
4.Literature Data Analysis on the Evolution Pattern of Traditional Chinese Medicine Syndromes in Psoriasis Vulgaris
Kewen GUAN ; Xiuli XIE ; Chuanjian LU
Journal of Traditional Chinese Medicine 2025;66(8):834-840
ObjectiveTo explore the distribution and evolution patterns of traditional Chinese medicine (TCM) syndromes and syndrome elements in psoriasis vulgaris (PV). MethodsLiterature related to TCM syndromes of PV published in databases including China National Knowledge Infrastructure (CNKI), Wanfang Data Knowledge Service Platform (Wanfang), VIP Chinese Technology Periodical Database (VIP), Chinese Biomedical Literature Database (CBM), PubMed, Embase, and Web of Science from their inception to December 31, 2023, was retrieved. Statistical analysis was conducted on the distribution of TCM syndromes and syndrome elements in the included studies. The data were further categorized into five-year periods to analyze the dynamic changes in syndromes and syndrome elements over time. ResultsA total of 2,853 studies were included, with 5,896 syndrome occurrences. The most common TCM syndromes in PV were blood-heat syndrome (2,167 occurrences, 36.75%), blood-stasis syndrome (1,219 occurrences, 20.68%), blood-dryness syndrome (1,124 occurrences, 19.06%), and damp-heat syndrome (263 occurrences, 4.46%). The most frequent syndrome categories included blood syndromes (4,680 occurrences, 79.38%) and dampness syndromes (347 occurrences, 5.89%). The most common syndrome elements related to disease location were blood division (4,874 occurrences, 94.38%) and spleen (99 occurrences, 1.92%). The most common syndrome elements related to disease nature were blood-heat (2,213 occurrences, 25.96%), blood-dryness (1,434 occurrences, 16.82%), and blood-stasis (1,330 occurrences, 15.60%). Except for the period 1978-1983, blood-heat, blood-stasis, and blood-dryness syndromes consistently ranked among the top three, with their combined proportion showing an overall upward trend (from 67.65% to 81.69%). The proportion of spleen deficiency with damp obstruction syndrome also increased (from 0.24% to 1.46%). In terms of syndrome classification, the proportion of blood syndromes showed an overall upward trend (from 67.65% to 83.46%), ranking first in all periods. The proportion of dampness syndromes showed a general downward trend (from 17.65% to 4.54%), ranking second after blood syndromes in most periods except for 1994-1998 and 1999-2003. The proportion of spleen-related syndromes showed an overall increase (from 0.24% to 1.85%). Regarding disease location elements, the proportion of blood division remained stable above 90%, while the proportion of spleen involvement increased (from 0.68% to 3.23%). As for disease nature elements, blood-heat (from 19.23% to 33.27%) and blood-stasis (from 1.92% to 20.83%) significantly increased, while dampness initially decreased and then slightly increased (from 11.54% to 5.73%). ConclusionIn the distribution of PV-related TCM syndromes, blood-heat, blood-stasis, and blood-dryness syndromes are the most common. Blood syndromes dominate syndrome classification, with disease location primarily in the blood division and disease nature characterized by blood-heat, blood-stasis, and blood-dryness. Evolutionary trends indicate that blood-heat, blood-stasis, and blood-dryness syndromes remain predominant and are increasing in proportion, while spleen deficiency with damp obstruction syndrome is also rising. Among syndrome classifications, the proportion of blood syndromes is increasing, dampness syndromes are decreasing, and spleen-related syndromes are on the rise. In terms of syndrome elements, blood division remains dominant, while spleen involvement is increasing. The proportion of blood-heat and blood-stasis is significantly increasing, while dampness first declines and then slightly rebounds. Overall, the mainstream TCM perspective of treating PV based on blood differentiation remains unchanged, with syndrome distribution focusing on blood division. The increasing importance of spleen deficiency and dampness in disease pathogenesis represents a new trend.
5.Association between lifestyle and cardiovascular-metabolic risk factor aggregation in a young and middle-aged male occupational population
Baoyi LIANG ; Lyurong LI ; Yingjun CHEN ; Lingxiang XIE ; Gaisheng LIU ; Liuquan JIANG ; Lu YU ; Qingsong CHEN
Journal of Environmental and Occupational Medicine 2025;42(4):385-391
Background Unhealthy lifestyle behaviors may be associated with an increased risk of cardiometabolic risk factor aggregation (CMRF≥ 2), and few studies have focused on the correlation between the two in occupational populations. Objective To investigate the current status of CMRF≥2 and the compliance of healthy lifestyle in male occupational personnel, explore the effect of lifestyle on cardiometabolic risk, and provide reference for formulating healthy behavior promotion strategies and reducing cardiometabolic risk in occupational populations. Methods The study subjects were selected from male workers who completed occupational health examinations at an occupational disease prevention and control hospital in Shanxi Province from May to December 2023, and
6.Effect of sodium-glucose cotransporter 2 inhibitor empagliflozin in alleviating uremic cardiomyopathy and related mechanism
Shi CHENG ; Yeqing XIE ; Wei LU ; Jiarui XU ; Yong YU ; Ruizhen CHEN ; Bo SHEN ; Xiaoqiang DING
Chinese Journal of Clinical Medicine 2025;32(2):248-258
Objective To investigate the effect of sodium-glucose cotransporter 2 inhibitor (empagliflozin, EMPA) on myocardial remodeling in a mouse uremic cardiomyopathy (UCM) model induced by 5/6 nephrectomy, through the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (PKB/AKT)/p65 signaling pathway. Methods The animals were divided into three groups: Sham group (n=6), UCM group (n=8), and UCM+EMPA group (n=8). A UCM model was established in C57BL/6N mice using the 5/6 nephrectomy. Starting from 5 weeks post-surgery, EMPA or a placebo was administered. After 16 weeks, blood pressure, serum creatinine, blood urea nitrogen, 24-hour urine glucose and urine sodium were measured. Cardiac structure and function were assessed by echocardiography. Hematoxylin-eosin (HE) staining and Masson trichrome staining were used to observe pathological changes in the heart and kidneys. Wheat germ agglutinin (WGA) staining was used to evaluate myocardial hypertrophy. The real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of myocardial hypertrophy- and fibrosis-related mRNAs. Western blotting was used to detect the expression levels of PI3K, AKT and p65 in myocardial tissues. Results After 16 weeks, UCM group exhibited significantly higher blood pressure, serum creatinine, blood urea nitrogen than sham group (P<0.01); UCM+EMPA group exhibited lower blood pressure, serum creatinine, blood urea nitrogen, and higher 24 h urine sodium and glucose than UCM group (P<0.05). Echocardiographic results showed ventricular remodeling in the UCM group, evidenced by left ventricular wall thickening, left ventricular enlargement, increased left ventricular mass, and decreased systolic function (P<0.05); ventricular remodeling was alleviated (P<0.05), though there was no significant improvement in systolic function in UCM+EMPA group. HE and Masson stainings revealed myocardial degeneration, necrosis, and interstitial fibrosis in UCM group (P<0.01); the myocardial pathology improved with reduced collagen deposition in UCM+EMPA group (P<0.01). WGA staining confirmed myocardial hypertrophy in UCM group (P<0.01), while myocardial hypertrophy was alleviated in UCM+EMPA group (P<0.01). RT-qPCR results showed myocardial hypertrophy- and fibrosis-related genes (NPPA, NPPB, MYH7, COL1A1, COL3A1, TGF-β1) were upregulated in UCM group (P<0.05), but downregulated in UCM+EMPA group. Western blotting showed PI3K, p-AKT/AKT ratio, and p-p65/p65 ratio were increased in UCM group, but decreased in UCM+EMPA group (P<0.05). Conclusion EMPA can improve myocardial hypertrophy and fibrosis in the UCM mouse model, and it may play the role through inhibiting the PI3K/AKT/p65 signaling pathway.
7.Treatment of pulmonary diseases in children from the lung collaterals′ structure, function and pathogenesis
Zhiyuan LU ; Yuhan WANG ; Qigang DAI ; Lili LIN ; Tong XIE ; Shouchuan WANG
Journal of Beijing University of Traditional Chinese Medicine 2025;48(3):323-329
The lung collaterals form a network that branches from the lung meridian, traversing the lung system and extending across the body′s surface. Lung collateral disease refers to the structural alterations or dysfunction in these collaterals caused by external or internal pathogens. Research into the structural and physiological functions of children′s lung collaterals, as well as the pathogenesis and syndrome differentiation for treating lung collateral diseases in children, holds significant value in guiding the prevention and treatment of pediatric respiratory conditions. Drawing on the theory of collateral disease, the clinical insights of both historical and contemporary physicians, and modern research findings—while considering the unique physiological and pathological characteristics of children′s respiratory systems—this study provides a foundational summary of the morphology and spatial distribution of children′s lung collaterals. The characteristics of these collaterals are highlighted as thin, sparse, short, narrow, brittle, and tender. From this structural understanding, the unique physiological functions of children′s lung collaterals are analyzed. The study further explores the interactions between pathogenic factors and lung collaterals, elucidating the pathogenesis and progression of children′s lung collateral diseases. It proposes treatment principles centered on "seeking treatment in the collaterals and employing the method of unblocking collaterals, "which align with the unique features of pediatric lung collaterals. Common treatment approaches, and relevant prescriptions for managing these diseases are summarized. This paper lays the foundation for a theoretical system encompassing the structure, function, pathogenesis, and syndrome differentiation for treating children′s lung collateral diseases. It offers valuable insights for the clinical diagnosis and management of pediatric respiratory diseases linked to collateral dysfunction and serves as a reference for the systematic development of a broader theoretical framework for children′s collateral diseases.
8.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
9.Shaoyaotang Alleviates Damage of Tight Junction Proteins in Caco-2 Cell Model of Inflammation by Regulating RhoA/ROCK Pathway
Nianjia XIE ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Yuting YANG ; Bo ZOU ; Da ZHAO ; Yi LU ; Mingsheng WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):70-77
ObjectiveTo investigate the protective effect and mechanism of Shaoyaotang (SYD) on the lipopolysaccharide (LPS)-induced damage of tight junction proteins in the human colorectal adenocarcinoma (Caco-2) cell model of inflammation via the Ras homolog gene family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway. MethodsCaco-2 cells were grouped as follows: Blank, model (LPS, 10 mg·L-1), SYD-containing serum (10%, 15%, and 20%), and inhibitor (Fasudil, 25 μmol·L-1). After 24 hours of intervention, the cell viability in each group was examined by the cell-counting kit 8 (CCK-8) method. Enzyme-linked immunosorbent assay was employed to determine the levels of endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the mRNA and protein levels, respectively, of RhoA, ROCK2, claudin-5, and zonula occludens-1 (ZO-1) in cells of each group. ResultsCompared with the blank group, the model group showcased a marked reduction in the cell viability (P<0.01), elevations in the levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), declines in both mRNA and protein levels of ZO-1 and claudin-5 (P<0.01), and rises in mRNA and protein levels of RhoA and ROCK2 (P<0.01). Compared with the model group, the Shaoyaotang-containing serum (10%, 15%, and 20%) groups had enhanced cell viability (P<0.01), lowered levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.01), up-regulated mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and down-regulated mRNA and protein levels of RhoA and ROCK2 (P<0.01). Moreover, the inhibitor group and the 15% and 20% Shaoyaotang-containing serum groups had lower levels of ET-1, TNF-α, IL-1β, and IL-6 (P<0.05, P<0.01), higher mRNA and protein levels of ZO-1 and claudin-5 (P<0.05, P<0.01), and lower mRNA and protein levels of RhoA and ROCK2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can lower the levels of LPS-induced increases in levels of inflammatory cytokines and endothelin to ameliorate the damage of tight junction proteins of the Caco-2 cell model of inflammation by regulating the expression of proteins in the RhoA/ROCK pathway.
10.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.


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