1.Expert Consensus on Clinical Application of Qidong Yixin Oral Liquid
Changkuan FU ; Xiaochang MA ; Mingjun ZHU ; Yue DENG ; Hongxu LIU ; Mingxue ZHANG ; Ying CHEN ; Yan ZHOU ; Ling ZHANG ; Jianhua FU ; Wei YANG ; Yu'er HU ; Ming CHEN ; Yanming XIE ; Yuanyuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):147-158
The prescription of Qidong Yixin oral liquid is derived from the experience of national medical master Ren Jixue in treating viral myocarditis (VMC). It has the functions of tonifying Qi, nourishing the heart,calming the mind, and relieving palpitations. It is used to treat VMC and angina pectoris of coronary heart disease caused by deficiency of both Qi and Yin. However,the understanding of its efficacy evidence, advantageous aspects, dosage and administration, and medication safety remains insufficient in clinical practice. Therefore,the development of the Expert Consensus on the Clinical Application of Qidong Yixin Oral Liquid (hereinafter referred to as consensus) was initiated. Consensus strictly followed the process and methods of the expert consensus on the clinical application of Chinese patent medicines of the China Association of Chinese Medicine,successively completing multiple tasks such as the consensus project initiation,determination of clinical problems,evidence search and evaluation,formation of recommendation opinions and consensus suggestions,solicitation of opinions,peer review, submission for review and release, and so on. Consensus formed a total of 10 recommendation opinions and 12 consensus suggestions,clarifying the clinical positioning,efficacy advantages,syndrome differentiation,dosage and administration,combination therapy,timing of medication,adverse reactions,contraindications, and precautions of Qidong Yixin oral liquid,indicating that it has good clinical advantages and safety in the treatment of VMC and angina pectoris of coronary heart disease,providing norms and references for physicians to safely and rationally apply Qidong Yixin oral liquid. Consensus was reviewed and approved for release by the Standardization Office of the China Association of Chinese Medicine on December 23, 2024. Standard number:GSCACM-376-2024.
2.Disease burden and changing trend in tracheal, bronchus, and lung cancer attributable to air pollution globally and in China and the United States from 1990 to 2021
Shoucai HU ; Chenglong YANG ; Lingling ZHANG ; Fu LI ; Yanan ZHANG ; Bin LIU ; Qingxin LI
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(01):97-104
Objective To systematically analyze the spatiotemporal distribution characteristics and epidemiological trends of tracheal, bronchus, and lung cancer (TBL) disease burden attributed to air pollution globally and in China and the United States from 1990 to 2021, and to assess the patterns of disease burden changes from 2022 to 2031 based on predictive models, providing a scientific basis for formulating targeted TBL prevention and control strategies. Methods Based on the Global Burden of Disease (GBD) 2021 database, we analyzed the disease burden data of TBL attributed to air pollution globally and in China and the United States from 1990 to 2021. R Studio 4.3.2 software was used to analyze the corresponding trends and the Bayesian age-period-cohort (BAPC) prediction model was used to predict the status of the disease burden of TBL attributed to air pollution in the world and in China and the United States from 2022 to 2031. Results In 2021, China had the highest number of deaths and disability-adjusted life years attributed to air pollution (211 400 patients and 4.8947 million person-years), followed by the United States (6 000 patients and 124 300 person-years). The age-standardized mortality rate (ASMR) and age-standardized disability-adjusted life years rate (ASDR) of TBL due to air pollution in the world and in China and the United States showed a decreasing trend. From 1990 to 2021, the ASMR and ASDR of TBL in China due to air pollution were much higher than those in the United States and the global average. In terms of gender, from 1990 to 2021, the disease burden of male patients with TBL attributed to air pollution was much higher than that of female patients. The BAPC prediction model showed that from 2022 to 2031, the ASMR and ASDR of TBL attributed to air pollution showed an upward trend globally, while they showed a downward trend in China and the United States. Conclusion Over the past 30 years, the air pollution-related TBL disease burden in the world and in China and the United States has continued to decline, but China's disease burden is still significantly higher than the global average. The disease burden in men far exceeds that in women, with men and the population aged ≥50 years being high-risk groups. In the future, the global disease trend may reverse and rise, while China and the United States are expected to continuously decline. However, precise prevention and control for high-risk groups remains a key challenge.
3.Compound Glycyrrhizin Tablets Ameliorate Liver Injury Induced by Tripterygium Glycosides Tablet by Regulating Cholesterol Metabolism
Xiaotong FU ; Chunyu CAO ; Chun LI ; Chenna LU ; Ting LIU ; Yifei YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):46-55
ObjectiveTo investigate the mechanism of liver injury induced by tripterygium glycosides tablets (TG) and the molecular mechanism of compound glycyrrhizin tablets (CG) in alleviating the abnormalities of cholesterol metabolism caused by TG via cholesterol metabolism. MethodsAccording to the body weights, male Sprague-Dawley (SD) rats were randomly grouped as follows: control (pure water), low-dose TG (TG-L, 189.0 mg·kg-1·d-1), high-dose TG (TG-H, 472.5 mg·kg-1·d-1), TG-L+CG (189.0 mg·kg-1·d-1 TG + 20.25 mg·kg-1·d-1 CG), and TG-H+CG (472.5 mg·kg-1·d-1 TG + 20.25 mg·kg-1·d-1 CG), with 6 rats in each group. Rats were administrated with corresponding drugs once daily for 3 weeks. At the end of the last administration, the mRNA and protein levels of liver X receptor-alpha (LXR-α), low-density lipoprotein receptor (LDLR), adenosine triphosphate-binding cassette transporter A1 (ABCA1), adenosine triphosphate-binding cassette transporter G1 (ABCG1), 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR), cholesterol 7α-hydroxylase (CYP7A1), cholesterol 12α-hydroxylase (CYP8B1), and sterol 27-hydroxylase (CYP27A1) in the liver tissue were determined by Real-time PCR and Western blotting, respectively. The level of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoAR), a regulatory enzyme of cholesterol synthesis, was measured by enzyme-linked immunosorbent assay (ELISA). HepG2 cells were used to observe the effect of TG on the cell proliferation in vitro. Specifically, HepG2 cells were grouped as follows: Low-dose TG (TG-l, 15 mg·L-1), medium-dose TG (TG-m, 45 mg·L-1), high-dose TG (TG-h, 135 mg·L-1), fenofibrate (FB, 10 μmol·L-1), CG extract, TG-h+FB (135 mg·L-1 TG + 10 μmol·L-1 FB), TG-m+FB (45 mg·L-1 TG + 10 μmol·L-1 FB), TG-l+FB (15 mg·L-1 TG + 10 μmol·L-1 FB), TG-h+CG (135 mg·L-1 TG + 60 μmol·L-1 CG), TG-m+CG (45 mg·L-1 TG + 60 μmol·L-1 CG), and TG-l+CG (15 mg·L-1 TG + 60 μmol·L-1 CG). The mRNA and protein levels of LXR-α, ABCG1, LDLR, CYP7A1, CYP8B1, and CYP27A1 in HepG2 cells were determined by Real-time PCR and Western blotting, respectively. ResultsThe rat experiment showed that compared with the control group, the TG-H group showed down-regulated mRNA levels of CYP7A1, CYP8B1, and CYP27A1 in the liver tissue (P<0.05, P<0.01), which were up-regulated by the application of CG (P<0.05, P<0.01), and the TG-H+CG group showed up-regulated mRNA level of LDLR (P<0.01). Compared with the control group, the TG-L and TG-H groups showed down-regulated protein levels of LDLR, CYP7A1, and CYP8B1 in the liver tissue (P<0.05, P<0.01). In addition, the protein levels of ABCG1 and LXR-α were down-regulated in the TG-H and TG-L groups, respectively (P<0.05). Compared with TG alone, TG+CG up-regulated the protein levels of ABCG1 and LDLR (P<0.05, P<0.01), and the protein levels of CYP7A1 and CYP8B1 in the TG-H+CG group were up-regulated (P<0.05, P<0.01). The cell experiment showed that compared with the control group, the TG-h group presented up-regulated mRNA level of LXR-α (P<0.01), and the TG-m and TG-h groups showcased down-regulated mRNA levels of LDLR and CYP7A1 (P<0.01) and up-regulated mRNA level of CYP27A1 (P<0.01) in HepG2 cells. The combination of CG with TG restored the above changes (P<0.01). Western blotting results showed that compared with the control group, the TG-m and TG-h groups showed down-regulated protein levels of LXR-α, ABCG1, LDLR, CYP7A1, CYP8B1, and CYP27A1 in HepG2 cells (P<0.01). Compared with the TG-h group, the TG-h+CG group showed up-regulated protein level of LDLR (P<0.05). Compared with the TG-m group, the TG-m+CG group showcased up-regulated protein levels of LDLR, ABCG1, CYP7A1, and CYP27A1 (P<0.05, P<0.01). ConclusionThe administration of TG at 189.0, 472.5 mg·kg-1 for 3 weeks could modulate the signaling pathways associated with cholesterol efflux, endocytosis, and cholesterol biotransformation in hepatocytes, leading to the accumulation of cholesterol and subsequent liver injury in rats. CG could ameliorate the liver injury induced by lipid metabolism disorders caused by TG by up-regulating the expression of LXR-α, LDLR, ABCG1, CYP7A1, CYP8B1, and CYP27A1 to promote cholesterol biotransformation.
4.Proteomics and Network Pharmacology Reveal Mechanism of Xiaoer Huatan Zhike Granules in Treating Allergic Cough
Youqi DU ; Yini XU ; Jiajia LIAO ; Chaowen LONG ; Shidie TAI ; Youwen DU ; Song LI ; Shiquan GAN ; Xiangchun SHEN ; Ling TAO ; Shuying YANG ; Lingyun FU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):69-79
ObjectiveTo explore the pharmacological mechanism involved in the treatment of allergic cough (AC) by Xiaoer Huatan Zhike granules (XEHT) based on proteomics and network pharmacology. MethodsAfter sensitization by intraperitoneal injection of 1 mL suspension containing 2 mg ovalbumin (OVA) and 100 mg aluminum hydroxide, a guinea pig model of allergic cough was constructed by nebulization with 1% OVA. The modeled guinea pigs were randomized into the model, low-, medium- and high-dose (1, 5, 20 g·kg-1, respectively) XEHT, and sodium montelukast (1 mg·kg-1) groups (n=6), and another 6 guinea pigs were selected as the blank group. The guinea pigs in drug administration groups were administrated with the corresponding drugs by gavage, and those in the blank and model groups received the same volume of normal saline by gavage, 1 time·d-1. After 10 consecutive days of drug administration, the guinea pigs were stimulated by 1% OVA nebulization, and the coughs were observed. The pathological changes in the lung tissue were observed by hematoxylin-eosin staining. The enzyme-linked immunosorbent assay was performed to measure the levels of C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), and malondialdehyde (MDA) in the bronchoalveolar lavage fluid (BALF) and immunoglobulin G (IgG) and immunoglobulin A (IgA) in the serum. Immunohistochemistry (IHC) was employed to observe the expression of IL-6 and TNF-α in the lung tissue. Transmission electron microscopy was employed observe the alveolar type Ⅱ epithelial cell ultrastructure. Real-time PCR was employed to determine the mRNA levels of IL-6, interleukin-1β (IL-1β), and TNF-α in the lung tissue. Label-free proteomics was used to detect the differential proteins among groups. Network pharmacology was used to predict the targets of XEHT in treating AC. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to search for the same pathways from the results of proteomics and network pharmacology. ResultsCompared with the blank group, the model group showed increased coughs (P<0.01), elevated levels of CRP, TNF-α, IL-6, and MDA and lowered level of SOD in the BALF (P<0.05, P<0.01), elevated levels of IgA and IgG in the serum (P<0.05, P<0.01), congestion of the lung tissue and infiltration of inflammatory cells, increased expression of IL-6 and TNF-α (P<0.01), large areas of low electron density edema in type Ⅱ epithelial cells, obvious swelling and vacuolization of the organelles, karyopyknosis or sparse and dissolved chromatin, and up-regulated mRNA levels of IL-6, IL-1β, and TNF-α (P<0.01). Compared with the model group, the drug administration groups showed reduced coughs (P<0.01), lowered levels of CRP, TNF-α, IL-6, and MDA and elevated level of SOD in the BALF (P<0.05, P<0.01), alleviated lung tissue congestion, inflammatory cell infiltration, and type Ⅱ epithelial cell injury, and decreased expression of IL-6 and TNF-α (P<0.01). In addition, the medium-dose XEHT group and the montelukast sodium group showcased lowered serum levels of IgA and IgG (P<0.05, P<0.01). The medium- and high-dose XEHT groups and the montelukast sodium showed down-regulated mRNA levels of IL-6, IL-1β, and TNF-α and the low-dose XEHT group showed down-regulated mRNA levels of IL-6 and TNF-α (P<0.05, P<0.01). Phospholipase D, mammalian target of rapamycin (mTOR), and epidermal growth factor receptor family of receptor tyrosine kinase (ErbB) signaling pathways were the common pathways predicted by both proteomics and network pharmacology. ConclusionProteomics combined with network pharmacology reveal that XEHT can ameliorate AC by regulating the phospholipase D, mTOR, and ErbB signaling pathways.
5.Nutritional supply status of school meals for primary and secondary students in three cities of Henan Province
LI Shan, YANG Li, HE Qidong, CAO Linlin, CHEN Xiaolong, LIU Bingrui, FENG Yinhua, FU Pengyu
Chinese Journal of School Health 2025;46(1):50-52
Objective:
To assess the nutritional supply status of school meals for primary and secondary school students in Henan Province, so as to provide a basis for scientific guidance of school meals.
Methods:
During 2021-2023, 115 lunch and dinner samples were collected from 25 primary and secondary schools in Zhoukou, Anyang and Luoyang of Henan Province by a direct selection method, and 13 nutrients were determined for each sample. The nutrient supply was evaluated based on Nutrition Guidelines of School Meals and Reference Intake of Dietary Nutrients for Chinese Residents (2023 Edition). Mann-Whitney U test was used to compare the differences of nutritional supply between urban and rural schools.
Results:
The median values for energy (709.77 kcal,1 kcal=4.18 kJ), fat energy supply ratio (0.27) and carbohydrate energy supply ratio (0.55) in the 66 lunches and dinners from primary school were within the recommended range. The supply of protein (28.39 g) and sodium (1 464.59 mg) was excessive. The median values of zinc (2.62 mg) and dietary fiber (5.19 g) were lower than the reference values. No statistically significant differences were observed in the supply of 13 nutrients between urban and rural primary schools( U = 427.00 -633.00, P > 0.05 ). Among 49 samples from secondary schools, the median value of energy supply (930.02 kcal), carbohydrate energy ratio ( 0.54 ) and fat energy supply ratio(0.25) were within the recommended range; and the median values of protein (38.82 g) and sodium (2 556.80 mg) were higher than the standard; and the median values of calcium (250.32 mg) and vitamin B1 (0.16 mg) were lower than the standard. Additionally, the differences in the level of vitamin B2 ( U =372.00) and zinc ( U =375.00) between the urban and rural secondary schools were statistically significant ( P <0.05).
Conclusion
Nutrient supply of primary and secondary school meals in three cities of Henan Province is inadequate and imbalanced, and the recipe need to be further optimized and improved.
6.Differential Analysis on Chemical Composition and Pharmacodynamic Effect Between Combined Decoction and Single Decoction of Famous Classical Formula Huaganjian
Yang WANG ; Gaoju ZHANG ; Ling LI ; Liping CHEN ; Li ZHANG ; Xiao LIU ; Yuyu ZHANG ; Yuan CUI ; Minglong LI ; Chaomei FU ; Xin YAN ; Yuxin HE ; Qin DONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):199-207
ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy, to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian, and meanwhile, the contents of four index components(geniposide, paeoniflorin, hesperidin and paeonol) were quantitatively analyzed by high performance liquid chromatography(HPLC). Nonalcoholic fatty liver disease(NAFLD) rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group, model group, lovastatin group(1.8 mg·kg-1), combined decoction group(1.26 g·kg-1) and single decoction group(1.18 g·kg-1). After successful modeling, lovastatin group, combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day, and the control group and model group were given equal amounts of normal saline by intragastric administration, after 4 weeks of administration, the serum and liver tissues were collected, and the contents of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in serum of rats were detected, and the liver pathological examination was carried out by hematoxylin-eosin(HE) staining and oil red O staining, so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian, and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide, paeoniflorin, hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples(P<0.01). The pharmacodynamic results showed that compared with the model group, both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats, reduce the serum levels of AST, ALT, TC, TG and LDL-C, increase the serum level of HDL-C, and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However, there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition, but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.
7.WANG Xiuxia's Clinical Experience in Treating Hyperprolactinemia with Liver Soothing Therapy
Yu WANG ; Danni DING ; Yuehui ZHANG ; Songli HAO ; Meiyu YAO ; Ying GUO ; Yang FU ; Ying SHEN ; Jia LI ; Fangyuan LIU ; Fengjuan HAN
Journal of Traditional Chinese Medicine 2025;66(14):1428-1432
This paper summarizes Professor WANG Xiuxia's clinical experience in treating hyperprolactinemia using the liver soothing therapy. Professor WANG identifies liver qi stagnation and rebellious chong qi (冲气) as the core pathomechanisms of hyperprolactinemia. Furthermore, liver qi stagnation may transform into fire or lead to pathological changes such as spleen deficiency with phlegm obstruction or kidney deficiency with essence depletion. The treatment strategy centers on soothing the liver, with a modified version of Qinggan Jieyu Decoction (清肝解郁汤) as the base formula. Depending on different syndrome patterns such as liver stagnation transforming into fire, liver stagnation with spleen deficiency, or liver stagnation with kidney deficiency, heat clearing, spleen strengthening, or kidney tonifying herbs are added accordingly. In addition, three paired herb combinations are commonly used for symptom specific treatment, Danggui (Angelica sinensis) with Chuanxiong (Ligusticum chuanxiong), Zelan (Lycopus lucidus) with Yimucao (Leonurus japonicus) , and Jiegeng (Platycodon grandiflorus) with Zisu (Perilla frutescens).
8.Study on the correlation of peak blood concentrations of compound sulfamethoxazole and its metabolites with clinical efficacy and adverse reactions in critically ill patients
Xiangxiang FU ; Lili ZHONG ; Jiangfan GU ; Mengyu MEI ; Xinxin LI ; Yang DENG ; Min WANG
China Pharmacy 2025;36(14):1775-1780
OBJECTIVE To analyze the correlation of the peak blood concentration (cmax) of compound sulfamethoxazole (TMP/SMZ) and its metabolite N-acetyl sulfamethoxazole (NSMZ) with clinical efficacy and adverse reactions in critically ill patients. METHODS The data of critically ill patients treated with TMP/SMZ in various ICU of Hainan General Hospital from December 2023 to January 2025 were retrospectively collected. The patients were divided into success group and failure group based on the treatment outcome. Simple linear regression and Spearman correlation analysis were used to analyze the correlation of TMP cmax, SMZ cmax, and NSMZ cmax with clinical efficacy and adverse reactions. The receiver operating characteristic curve (ROC) was used to determine the cutoff values of cmax for predicting the occurrence of adverse reactions. RESULTS Among critically ill patients with an acute physiology and chronic health evaluation Ⅱ (APACHE-Ⅱ) ≥15 points 24 h of check-in at ICU, SMZ cmax of success group was significantly higher than failure group (P<0.05). The daily total dose of TMP/SMZ was positively correlated with TMP cmax and SMZ cmax( P<0.05). TMP cmax was significantly correlated with hepatotoxicity and nephrotoxicity, SMZ cmax with hepatotoxicity, and NSMZ cmax with nephrotoxicity (P<0.05). The cutoff values of TMP cmax for predicting nephrotoxicity and hepatotoxicity were 7.25 μg/mL and 6.63 μg/mL, respectively. The cutoff value of SMZ cmax for predicting hepatotoxicity was 138.00 μg/mL, and that of NSMZ cmax for predicting nephrotoxicity was 60.76 μg/mL. CONCLUSIONS Among critically ill patients with an APACHE-Ⅱ ≥15 points 24 h of check-in at ICU, SMZ cmax is associated with treatment success. Hepatotoxicity risk significantly increases when TMP cmax ≥6.63 μg/mL or SMZ cmax ≥138.00 μg/mL; nephrotoxicity risk significantly increases when TMP cmax ≥7.25 μg/mL or NSMZ cmax ≥60.76 μg/mL.
9.Preliminary study on the biological characteristics of heat shock cognate protein 20 of Schistosoma japonicum
Xingang YU ; Kaijian YUAN ; Yilong LI ; Xuanru MU ; Hui XU ; Qiaoyu LI ; Wenjing ZENG ; Zhiqiang FU ; Yang HONG
Chinese Journal of Schistosomiasis Control 2025;37(3):294-303
Objective To clone and express the heat shock cognate protein 20 (SjHsc20) of Schistosoma japonicum, and to preliminarily investigate its biological characteristics. Methods The target fragment of the SjHsc20 gene was amplified using PCR assay and cloned into the pET-28a(+) expression plasmid to generate the recombinant expression vector pET-28a(+)-SjH-sc20, which was then transformed into Escherichia coli BL21 (DE3) competent cells. The recombinant SjHsc20 (rSjHsc20) protein was induced with isopropyl β-D-thiogalactopyranoside (IPTG) and purified, and the expression of the rSjHsc20 protein was checked with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the rSjHsc20 protein was detected using Western blotting, and the transcriptional levels of SjHsc20 were quantified in S. japonicum worms at different developmental stages and in male and female adult worms using real-time quantitative PCR (RT-qPCR) assay. Thirty female BALB/c mice at ages 6 to 8 weeks were divided into three groups, including the rSjHsc20 immunization group, the PBS control group, and the ISA 206 adjuvant group, of 10 mice in each group. Mice in the rSjHsc20 immunization group were subcutaneously immunized with 20 μg rSjHsc20 on days 1, 15 and 31, and animals in the PBS control group were subcutaneously injected with the same volume of PBS on days 1, 15 and 31, while mice in the ISA 206 adjuvant group were subcutaneously immunized with the same volume of ISA 206 adjuvant on days 1, 15 and 31, respectively. All mice in each group were infected with (40 ± 2) S. japonicum cercariae via the abdomen 14 day following the last immunization. Levels of serum specific IgG and its subtypes IgG1 and IgG2 antibodies against rSjHsc20, and the serum titers of anti-rSjHsc20 antibody were detected in mice using indirect enzyme-linked immunosorbent assay (ELISA). All mice were sacrifice 42 days post-infection, and S. japonicum worms were collected from the hepatic portal vein and counted. The eggs per gram (EPG), worm burden reductions and egg burden reductions were estimated to evaluate the protective efficacy of the rSjHsc20 protein. Results The SjHsc20 gene had an open reading frame (ORF) with 756 bp in length and encoded 252 amino acids, and the rSjHsc20 protein had a relative molecular mass of approximately 29 kDa. The rSjHsc20 protein was recognized by the serum of mice infected with S. japonicum and the serum of mice immunized with the rSjHsc20 protein, indicating that rSjHsc20 had a good immunogenicity. There was a significant difference in the transcriptional levels of the SjHsc20 gene among the 7-day (1.001 4 ± 0.065 7), 12-day (2.268 3 ± 0.129 2), 21-day (1.378 5 ± 0.160 4), 28-day (1.196 4 ± 0.244 0), 35-day (1.646 3 ± 0.226 1), 42-day worms of S. japonicum (1.758 0 ± 0.611 1) (F = 38.45, P < 0.000 1), and the transcriptional level of the SjHsc20 gene was higher in the 12-day worms than in worms at other developmental stages (all P values < 0.000 1). The serum levels of anti-rSjHsc20 IgG antibody were 0.106 6 ± 0.010 7, 0.108 3 ± 0.010 4, and 0.553 2 ± 0.069 1 in the PBS control group, ISA 206 adjuvant group, and rSjHsc20 immunization group following the last immunization, respectively, and the serum levels of IgG1 antibody were 0.137 3 ± 0.054 0, 0.181 1 ± 0.096 8, and 1.765 8 ± 0.221 1, while the levels of IgG2a antibody were 0.280 3 ± 0.197 6, 0.274 0 ± 0.146 3, and 1.560 4 ± 0.106 0, respectively. There were significant differences in the serum levels of anti-rSjHsc20 IgG (F = 397.70, P < 0.000 1), IgG1 (F = 401.00, P < 0.000 1) and IgG2a antibodies (F = 229.70, P < 0.000 1) among the three groups, and the serum levels of anti-rSjHsc20 IgG, IgG1 and IgG2a antibodies were higher in the rSjHsc20 immunization group than in the PBS control group and the ISA 206 adjuvant group (all P values < 0.000 1). There was a significant difference in the IgG1/IgG2a ratio among the rSjHsc20 immunization group (1.177 2 ± 0.143 6), the PBS control group (0.428 4 ± 0.199 8) and the ISA 206 adjuvant group (0.559 9 ± 0.181 1) (F = 43.97, P < 0.000 1), and the IgG1/IgG2a ratio was > 1 in the rSjHsc20 immunization group, which was higher than in the PBS control group and the ISA 206 adjuvant group (both P values < 0.000 1). The titers of serum anti-rSjHsc20 antibody were all above 1∶16 384 in the rSjHsc20 immunization group following immunizations on days 1, 15 and 31, indicating that the rSjHsc20 protein had a strong immunogenicity. The mean worm burdens were (16.60±5.75), (15.80±5.58) worms per mouse and (14.40±5.75) worms per mouse in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group 42 days post-infection with S. japonicum cercariae (F = 0.50, P > 0.05), and the EPG were 68 370 ± 22 690, 67 972 ± 19 502, and 41 075 ± 13 251 in the PBS control group, the ISA 206 adjuvant group and the rSjHsc20 immunization group (F = 4.55, P < 0.05), with lower EPG in the PBS control group and the ISA 206 adjuvant group than in the rSjHsc20 immunization group (both P values < 0.05). Immunization with the rSjHsc20 protein resulted in a worm burden reduction of 13.25% and an egg burden reduction of 39.92% relative to the PBS control group. Conclusions SjHsc20 is successfully cloned and expressed, and the rSjHsc20 protein induces partial immunoprotective effects in mice, which provides a basis for deciphering the biological functions of SjHsc20 and assessing the potential of SjH-sc20 as a vaccine candidate.
10.Effects of total extract of Anthriscus sylvestris on immune inflammation and thrombosis in rats with pulmonary arterial hypertension based on TGF-β1/Smad3 signaling pathway.
Ya-Juan ZHENG ; Pei-Pei YUAN ; Zhen-Kai ZHANG ; Yan-Ling LIU ; Sai-Fei LI ; Yuan RUAN ; Yi CHEN ; Yang FU ; Wei-Sheng FENG ; Xiao-Ke ZHENG
China Journal of Chinese Materia Medica 2025;50(9):2472-2483
This study aimed to explore the effects and mechanisms of total extracts from Anthriscus sylvestris on pulmonary hypertension in rats. Sixty male SD rats were divided into normal(NC) group, model(M) group, positive drug sildenafil(Y) group, low-dose A. sylvestris(ES-L) group, medium-dose A. sylvestris(ES-M) group, and high-dose A. sylvestris(ES-H) group. On day 1, rats were intraperitoneally injected with monocrotaline(60 mg·kg~(-1)) to induce pulmonary hypertension, and the rat model was established on day 28. From days 15 to 28, intragastric administration of the respective treatments was performed. After modeling and treatment, small animal echocardiography was used to detect the right heart function of the rats. Arterial blood gas was measured using a blood gas analyzer. Hematoxylin and eosin(HE) staining and Masson staining were performed to observe cardiopulmonary pathological damage. Flow cytometry was used to detect apoptosis in the lung and myocardial tissues and reactive oxygen species(ROS) levels. Western blot was applied to detect the expression levels of transforming growth factor-β1(TGF-β1), phosphorylated mothers against decapentaplegic homolog 3(p-Smad3), Smad3, tissue plasminogen activator(t-PA), and plasminogen activator inhibitor-1(PAI-1) in lung tissue. A blood routine analyzer was used to measure inflammatory immune cell levels in the blood. Enzyme-linked immunosorbent assay(ELISA) was used to detect the expression levels of P-selectin and thromboxane A2(TXA2) in plasma. The results showed that, compared with the NC group, right heart hypertrophy index, right ventricular free wall thickness, right heart internal diameter, partial carbon dioxide pressure(PaCO_2), apoptosis in cardiopulmonary tissue, and ROS levels were significantly increased in the M group. In contrast, the ratio of pulmonary blood flow acceleration time(PAT)/ejection time(PET), right cardiac output, change rate of right ventricular systolic area, systolic displacement of the tricuspid ring, oxygen partial pressure(PaO_2), and blood oxygen saturation(SaO_2) were significantly decreased in the M group. After administration of the total extract of A. sylvestris, right heart function and blood gas levels were significantly improved, while apoptosis in cardiopulmonary tissue and ROS levels significantly decreased. Further testing revealed that the total extract of A. sylvestris significantly decreased the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and PAI-1 proteins in lung tissue, while increasing the expression of t-PA. Additionally, the extract reduced the levels of inflammatory cells such as leukocytes, lymphocytes, granulocytes, and monocytes in the blood, as well as the levels of P-selectin and TXA2 in plasma. Metabolomics results showed that the total extract of A. sylvestris significantly affected metabolic pathways, including arginine biosynthesis, tyrosine metabolism, and taurine and hypotaurine metabolism. In conclusion, the total extract of A. sylvestris may exert an anti-pulmonary hypertension effect by inhibiting the TGF-β1/Smad3 signaling pathway, thereby alleviating immune-inflammatory responses and thrombosis.
Animals
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Male
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Smad3 Protein/metabolism*
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Transforming Growth Factor beta1/metabolism*
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Rats, Sprague-Dawley
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Rats
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Signal Transduction/drug effects*
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Hypertension, Pulmonary/genetics*
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Thrombosis/immunology*
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Drugs, Chinese Herbal/administration & dosage*
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Humans
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Apoptosis/drug effects*


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